JP2017536385A - Antifungal compound - Google Patents

Abstract

The present invention relates to compounds defined herein useful in the treatment of mycosis, compositions comprising them, and their use in therapy. [Selection figure] None

Description

(Field of Invention)
The present invention relates to compounds useful in the treatment of mycoses, compositions containing them, and their use in therapy.

(Background of the invention)
The incidence of fungal infections has increased significantly over the past 20 years, and invasive forms are a major cause of morbidity and mortality, especially in immunocompromised or immunosuppressed patients. Disseminated candidiasis, pulmonary aspergillosis, and emerging opportunistic fungi are the most common factors that cause these severe mycoses. It is a unique feature of fungi that fungi can generate extracellular matrix (ECM) that binds them and attaches them to their in vitro or in vivo substrates. These biofilms serve to protect the fungus from the unfavorable environment of the host immune system and to resist killing by antimicrobial agents (Kaur and Singh, 2013).

  Pulmonary aspergillosis can be divided into patients suffering from non-invasive diseases and patients with invasive conditions. A further subdivision is used to characterize patients who exhibit allergic components for aspergillosis (ABPA; known as allergic bronchopulmonary aspergillosis) compared to patients who do not exhibit allergic components for aspergillosis. Factors that promote pulmonary aspergillosis can be acute, such as exposure to high doses of immunosuppressive drugs or exposure to intubation in the intensive care unit. Alternatively, this factor may be chronic, such as infection with previous TB (Denning et al., 2011a). Chronic lung infection with Aspergillus can cause patients to have extensive and permanent lung injury and require lifelong treatment with oral azole drugs (Limper et al., 2011).

  Successive studies suggest that Aspergillus infection may play an important role in clinical asthma (Chishimba et al., 2012; Pasqualotto et al., 2009). In addition, recently published studies have linked Aspergillus infection to worse clinical outcomes in patients with COPD (Bafadhel et al., 2013). Similarly, cross-sectional studies have shown an association between the presence of Aspergillus and Candida species in sputum and worsening lung function (Chotirmall et al., 2010; Agbetile et al., 2012). .

  Invasive aspergillosis (IA) exhibits high mortality in immunocompromised patients, such as immunocompromised patients undergoing allogeneic stem cell transplantation or solid organ transplantation (eg lung transplantation). The first IA case reported in immunocompromised patients occurred in 1953. This event overlapped with the introduction of corticosteroids and cytotoxic chemotherapy into the treatment regimen (Rankin, 1953). Invasive aspergillosis is of great interest in the treatment of leukemia and other hematological malignancies given its high incidence and associated mortality. Mortality is usually over 50% (Lin et al., 2001) and long term rates are up to 90% in allogeneic hematopoietic stem cell transplant recipients despite the availability of oral triazole drugs (Salmeron et al., 2012). In patients undergoing solid organ transplantation (especially pulmonary), the use of high doses of steroids makes patients more susceptible to infection (Thompson and Patterson, 2008), which is a serious problem. This disease is also seen in a less severe population of immunocompromised patients. These include those suffering from causative COPD or cirrhosis, patients receiving high-dose steroids, and those who have central venous catheters or who are maintained on a ventilator ( Dimopoulos et al., 2012).

  Existing antifungal drugs are mainly administered orally or systemically. These commonly utilized routes of delivery are weak for the treatment of pulmonary respiratory tract infections because the drug concentration achieved at the site of infection tends to be lower than in the organ. This is especially true for the liver, which is a site of toxicity: up to 15% of patients treated with voriconazole suffer from elevated transaminase levels (Levin et al., 2007; Lat and Thompson, 2011). Liver exposure also results in significant drug interactions resulting from inhibition of liver P450 enzymes (Jeong et al., 2009; Wexler et al., 2004).

  In addition, the widespread use of triazoles in both hospitals and agriculture has led to an increasing incidence of problematic and resistant mycosis in several places (Denning et al., 2011b; Bowyer and Denning's Literature, 2014).

  Clearly, there is an urgent medical need for new antifungal agents that provide improved efficacy and a better systemic tolerability profile.

(式中:
Rは、水素、ハロ、シアノ、C_1-4ハロアルキル、C_1-4ハロアルコキシ、又はSO₂NR⁵R⁶を表し;
R^1a及びR^1bは、独立に、水素又はハロを表し;
R²は、水素、ハロ、シアノ、C_1-4アルキル、C_1-4アルコキシ、又はC_1-4ハロアルコキシを表し;
R³は、ハロ、シアノ、C_1-4アルキル、又はC_1-4ヒドロキシアルキルを表し;
R⁴は、水素又はC_1-4アルキルを表し;
Xは、CH又はNを表し;
Yは、CH又はNを表し;
R⁵及びR⁶は、独立に、水素又はC_1-4アルキルを表す。) (Summary of Invention)
In a first aspect, the present invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof (hereinafter sometimes referred to as “compounds of the invention” or “compounds of the present disclosure”).

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
X represents CH or N;
Y represents CH or N;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl. )

  The biological data disclosed hereinbelow reveals that the compounds of the present invention are potent inhibitors of Aspergillus fumigatus growth in an in vitro assay. In immunosuppressed mice, the compounds of the invention showed potent inhibition of Aspergillus fumigatus infection. Other desirable properties of the compounds of the present invention are described herein.

(Brief description of the drawings)
FIG. 1 shows the effect of prophylactic and therapeutic treatment with Compound Example 1 on CFU in the lungs of immunodeficient neutropenic mice infected with Aspergillus fumigatus. FIG. 2 shows the effect of prophylactic or therapeutic treatment with Compound Example 1 on the galactomannan concentration in BALF of immunodeficient neutropenic mice infected with Aspergillus fumigatus. " FIG. 3 shows the effect of prophylactic or therapeutic treatment with Compound Example 1 on the galactomannan concentration in the serum of immunodeficient neutropenic mice infected with Aspergillus fumigatus. 1-3, the symbol ^*** indicates significance of P <0.001.

(Detailed description of the invention)
The alkyl group can be branched or straight chain. A C _1-4 alkyl group may represent, for example, C _1-3 alkyl or C _1-2 alkyl. Exemplary alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, and CH ₂ CHMe ₂ . In one embodiment, alkyl refers to straight chain alkyl.

  Alkoxy as used herein means —Oalkyl and includes straight or branched chain alkoxy such as methoxy, ethoxy, propoxy, butoxy.

  Hydroxyalkyl means an alkyl having a hydroxyl substituent at any position. Examples include hydroxymethyl, 2-hydroxyethyl, 3-hydroxy-n-propyl, and 4-hydroxy-n-butyl.

  The halogen may suitably be Br, Cl or F, especially Cl or F, especially F.

  In one embodiment, pharmaceutically acceptable salts of the compounds of the invention are provided.

  The compounds of the present disclosure include compounds where the specified atom is a natural or non-natural isotope. In one embodiment, the isotope is a stable isotope. Accordingly, the compounds of the present disclosure include, for example, compounds containing one or more deuterium atoms instead of hydrogen atoms.

  The present disclosure also extends to all polymorphic forms of the compounds defined herein.

  The present disclosure also extends to all solvates of the compounds defined herein. Examples of solvates include hydrates.

Suitably, R represents, H, F, CN, OCHF 2, OCF 3, SO 2 NH 2, SO 2 NHMe, SO 2 NMe 2, or Cl, especially, F or CN, more preferably represents F.

Suitably R ^1a represents H, F or Cl, in particular H.

Suitably R ^1b represents H or F, especially H.

Suitably R ² represents H, Me or OMe, especially H.

Suitably R ³ represents Me, CN, Cl, CH ₂ OH or F, especially Me.

Suitably R ⁴ represents H or Me, in particular H.

Suitably R ⁵ represents H or Me.

Suitably R ⁶ represents H or Me.

Suitably NR ⁵ R ⁶ represents NH ₂ , NHMe or NMe ₂ .

  Suitably X represents CH or N, especially CH.

Suitably Y represents CH (this carbon atom may optionally be substituted by R, R ^1a or R ^1b , eg by F) or N, especially CH.

Suitably R ^1a and R ^1b each represent H and R represents F, CN, OCHF ₂ , OCF ₃ , SO ₂ NH ₂ , SO ₂ NHMe, SO ₂ NMe ₂ or Cl, especially , R ^1a and R ^1b represent H, R represents F or CN, more preferably R ^1a and R ^1b represent H and R represents F.

Alternatively, preferably R ^1a represents F, R ^1b represents H, R represents F or OCHF ₂ , for example, R ^1a represents F, R ^1b represents H, and R represents F Represent.

Alternatively, preferably R ^1a represents Cl, R ^1b represents H and R represents F.

Suitably R ⁴ represents H, R ³ represents Me, CN, Cl, CH ₂ OH or F, in particular R ⁴ represents H and R ³ represents Me.

Alternatively, preferably R ⁴ represents Me and R ³ represents Me.

Suitably Y represents CH, R ^1a and R ^1b each represent H and R is in the 3- or 4-position, especially the 4-position.

Alternatively, preferably Y represents CH, R ^1b represents H and R and R ^1a are in the 2,4, 3,4, 2,5, or 3,5 positions, especially 2, 4th.

Alternatively, preferably Y represents N, R ^1a and R ^1b each represent H, and R is in the 4-, 5-, or 6-position, especially the 5-position.

Suitably Y represents CH and R, R ^1a and R ^1b are in the 2, 4, and 6 positions.

Suitably R ² is ortho to the nitrogen of the piperazinyl substituent. Alternatively, R ² is meta to the nitrogen of the piperazinyl substituent.

Suitably R ⁴ is ortho to the oxygen of the ether substituent.

  Preferably, the aromatic moiety comprising Y is 4-fluorophenyl, 3-fluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 4-cyanophenyl, 3-cyanophenyl, 4-difluoromethoxyphenyl, 4- (trifluoromethoxy) phenyl, 4-sulfamoylphenyl, 4- (N-methylsulfamoyl) phenyl, 4- (NN-dimethyl Sulfamoyl) phenyl, 4-cyano-2-fluorophenyl, 4- (difluoromethoxy) -3-fluorophenyl, 4-chloro-2-fluorophenyl, 4-chloro-3-fluorophenyl, 2,4,6 -Trifluorophenyl, 5-fluoropyridin-2-yl, 5-cyanopyridin-2-yl, 6-cyanopyridin-2-yl, 4-cyanopyridin-2-yl, 5- (trifluoromethoxy) pyridine- 2-yl or 5-chloropyridin-2-yl, and Divided, 4-fluorophenyl or 4-cyanophenyl, more preferably represents 4-fluorophenyl.

In one embodiment, the compound of formula (I) is:
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (2,4-difluorophenyl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-cyanophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -2,5-dimethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (3-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-chlorophenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3,5-dimethylphenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-difluoromethoxyphenyl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (trifluoromethoxy) phenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3,5-dimethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-hydroxymethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-chlorophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-sulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (N-methylsulfamoyl) phenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (4- (N, N-dimethylsulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (4-sulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyano-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-fluorophenyl) piperazin-1-yl) -N- (4-cyano-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (difluoromethoxy) -3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (2,5-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (3,4-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (3,5-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-chloro-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-chloro-3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (2,4,6-trifluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -3-methylbenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -2-methylbenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -3-methoxybenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -2-methoxybenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (6-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-hydroxymethylphenyl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5- (trifluoromethoxy) pyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-chloropyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 6-methylpyridin-2-yl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide and their pharmaceutically acceptable salts.

In one embodiment, the compound of formula (I) is:
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) Methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide, or a pharmaceutically acceptable salt thereof.

The compounds of the present invention can be prepared from commercially available starting materials by the non-limiting synthetic methods shown below (Schemes 1-3).
Scheme 1

  Thus, the compound of formula (I) can be reacted with a benzoic acid precursor (II) or a suitably protected derivative thereof with an activator to produce a reactive electrophilic carboxylic acid derivative, after which Can be obtained by a general process (Scheme 1) in which it is reacted with an amine of formula (III) or a suitably protected derivative thereof. In some cases, activated carboxylic acid derivatives such as acid chlorides may be isolated, or in other cases, transiently produced in situ and used as such, rather than being isolated. It will be understood by those skilled in the art that these may be intermediates. Suitable reagents for activating the carboxylate group include carbonyldiimidazole, 1-chloro-N, N, 2-trimethylprop-1-en-1-amine, and a wide selection of peptide coupling agents such as And benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP®), N- (3-dimethylaminopropyl) -N′-ethylcarbodiimide hydrochloride (EDCI.HCl), and the like. Such a reaction is conveniently carried out in an apolar aprotic solvent such as DCM, or optionally in a polar aprotic solvent such as pyridine. The resulting amide may be produced at ambient temperatures such as RT or below, or may be formed at elevated temperatures, for example 60 ° C. A review of amide preparation methods is given in: “Amide bond formation and peptide coupling”, Montalbetti, CAGN and Falque, V., Tetrahedron, 2005, 61, 10827-10852. Yes. When one or more protecting groups are utilized by a suitable deprotection step, the compound of formula (I) appears.

The intermediate of formula (II) is first derived by a noble metal mediated bond formation process, such as the Buchwald coupling reaction, between the piperazine derivative of formula (V) and the 4-bromobenzoate of formula (VI). In addition, the corresponding benzoic acid ester of formula (IV) can be provided. One skilled in the art will appreciate that a wide variety of conditions can be used to affect this type of transformation. In particular, palladium catalysts and phosphine ligands such as RuPhosG3 and RuPhos are routinely utilized in the presence of a base such as cesium carbonate or lithium hexamethyldisilazane. Such coupling procedures are generally carried out in polar aprotic solvents such as DMF at elevated temperatures, for example 70-80 ° C. The compound of formula (II) is obtained after subsequent hydrolysis of the ester (IV) to the free acid. Suitable conditions for this functional group interconversion depend on the nature of the ester. Primary and secondary esters (e.g., R ^a = Me, Et, and Pr ⁱ ) are suitable inorganic bases in aqueous mixtures of aprotic and / or protic solvents such as THF: methanol: water, such as It is conveniently saponified by exposure to lithium hydroxide. More disturbing examples (eg R ^a = ^t Bu) are strong mineral acids, such as hydrochloric acid or strong organic acids, typically trifluoro, used neat or in the presence of a solvent such as DCM. It can be more easily deesterified by treatment with acetic acid.

In another process using the Buchwald coupling method, the advanced ester intermediate of formula (IV) is converted to an aryl bromide of formula (VIII) and a piperazine motif (R) of formula (XIV) in a manner similar to that described above. ^a = lower alkyl, eg C _1-5 alkyl, eg methyl or ethyl, or tert-butyl). The same technique can be applied to aryl bromides (VIII) and suitably protected piperazines of formula (IX), thereby producing intermediates of formula (VII). The intermediate is converted to the compound of formula (V) described above by a suitable deprotection step. A suitable nitrogen protecting group strategy for this purpose is, for example, a Boc group that is stable under the conditions required for this coupling reaction and can be removed later by treatment with acid (P = CO ₂ ^t Bu). It is a urethane that uses Removal of the N-Boc protecting group is usually accomplished at ambient temperature in an inert solvent such as DCM by treatment with a strong organic acid such as TFA.

In a third approach, the benzoate of formula (IV) is obtained by reaction of the phenolic intermediate (XIII) [Y = CH] or the corresponding pyridinol [Y = N] with an alkylating agent of formula (XI). Where Z represents a leaving group and the stoichiometric composition of the reagent is absolute as indicated. Typical alkylating agents for this type of transformation include sulfonate esters such as mesylate (Z = Me ₂ SO ₂ O) or triflate (Z = CF ₃ SO ₂ O) and alkyl halides such as Chloromethyl or bromo-methyl derivatives (Z = Cl and Br, respectively).

An important practical consideration is the tosylate derivative (XIa) [Z = p-tolyl SO ₂ O], ie: ((3S, 5R) -5-((1H-1,2,4-triazol-1-yl ) Methyl-5- (2,4-difluorophenyl) tetrahydrofuran-3-yl) methyl 4-methylbenzenesulfonate is a large, widely used reagent from commercial sources with high enantiomeric purity. This derivative is also chemically and configurationally stable and, as a solid, offers operational advantages over other more volatile (and therefore more dangerous) alkylating agents. The etherification process is usually carried out under basic conditions, for example in the presence of sodium ethoxide (to generate the phenoxide anion in situ) in a polar aprotic solvent, preferably in DMF. It is carried out at a temperature in the range of ~ 50 ° C.

In a manner similar to that already described above, the 1,4-diarylated piperazine of formula (XIII) is converted to the mono-substituted piperazine (XIV) and the bromophenol of formula (X) [Y = CH, R ^c = H Or the corresponding bromo-pyridinol [Y = N, R ^c = H] or, optionally, their Buchwald coupling with these suitably protected derivatives [R ^c ≠ H]. When a compound of formula (X) is (for example, as methyl or benzyl ether) is protected, the desired coupling product (XIII) is suitable deprotection step (e.g., by BBr ₃ O-demethylation or Obtained after hydrocracking).

  The hydroxylated aryl bromide of formula (X) is also converted to the N-aryl piperazine of formula (XII) in the same way by a coupling reaction with a monoprotected piperazine of formula (IX). A commonly used amine protecting group for such purposes is the Boc group, in which case the compound of formula (IX) is tert-butylpiperazine-1-carboxylate. It will be apparent to those skilled in the art that other nitrogen protecting groups can be selected based on considerations of orthogonality and operational efficiency. Alkylation of the free hydroxyl function in the N-aryl piperazine of formula (XII) with tosylate (XIa) as described above results in an intermediate of formula (VII).

  In some cases, the preparation outlined above may be advantageous to carry out the same or similar synthetic transformations in a different order in order to improve the overall efficiency of the process and / or the quality of the material obtained therefrom. It will be understood from the route (Scheme 1). In addition to the examples already disclosed, the hydroxylated aryl bromide of formula (X) is converted to the compound of formula (VII) by performing the two steps outlined above in reverse order. be able to. Treatment of phenol / pyridinol of formula (X) with tosylate (XIa) provides an ether derivative of formula (VIII). The ether derivative was converted to an intermediate of formula (VII) under Buchwald coupling conditions as described above.

Further strategies for preparing the compounds of the invention using the synthetic techniques described above are revealed below (Scheme 2), where the same or closely related intermediates found above (Scheme 1 ) Are assembled in a different order.
Scheme 2

  Treatment of aniline component (III) with benzoyl chloride (XIX) provides benzanilide derivatives of formula (XVIII). As already described, such amide products can be obtained directly from the corresponding amine and benzoic acid using a wide variety of activators, including peptide coupling reagents, many of which are utilized in the art. Can be prepared. These products are subjected to a Buchwald coupling reaction with a suitable mono-protected piperazine (IX) under the action of a catalyst in the manner described above to yield an intermediate of formula (XVII). Piperazine derivatives that are stable under these bond-forming conditions include urethanes such as N-Boc derivatives, but in some cases another (eg, Cbz group) may be advantageous. Removal of the amine protecting group under suitable conditions and a second palladium-mediated N-arylation procedure using an aromatic bromide of formula (X) or a protected derivative thereof yields an advanced intermediate of formula (XV). Supplied.

  In some cases, this conversion can be accomplished using a substrate (X) in which a free hydroxyl group is present (ie, Rc = H). In other cases it may be preferred to protect the functional group, for example as an ether derivative, typically as a benzyl ether. These can be converted back to free phenol or pyridinol by O-dealkylation under hydrocracking conditions. A representative compound of formula (I) is then obtained by subsequent alkylation of the hydroxyl group with tosylate (XIa) with a reagent of formula (XI).

A compound of the invention in which a benzamide is formed with 2-aminopyridine, (I) [Y = N] is a combination of a primary carboxamide of formula (XX) and an optionally substituted 2-bromopyridine substrate of formula (XXI) Available from palladium-mediated cross coupling (Scheme 3). Typical reaction conditions for effecting buchwald amidation of aryl halides are tris (dibenzylideneacetone) in the presence of phosphine ligands, generally Xantphos, for example, basic conditions with cesium carbonate. Including the use of palladium catalysts such as dipalladium (O). It is common to carry out such reactions in polar aprotic solvents, typically DMF, at elevated temperatures such as 80-100 ° C. The primary benzamide (XX) is easily obtained from the benzoic acid (II) by treating the corresponding benzoic acid (II) with a source of ammonia, conveniently ammonium chloride, under standard peptide coupling conditions. Generate.
Scheme 3

  Protecting groups and their means of removal are described in "Protective Groups in Organic Synthesis" by Theodora W. Greene and Peter GM Wuts, published by John Wiley &Sons; 4th edition, 2006, ISBN-10: It is described in 0471697540. A review of amide preparation methods is given in: “Amide bond formation and peptide coupling”, Montalbetti, CAGN and Falque, V., Tetrahedron, 2005, 61, 10827-10852. Yes.

(式中:
R²は、水素、ハロ、シアノ、C_1-4アルキル、C_1-4アルコキシ、又はC_1-4ハロアルコキシを表し;
R³は、ハロ、シアノ、C_1-4アルキル、又はC_1-4ヒドロキシアルキルを表し;
R⁴は、水素又はC_1-4アルキルを表し;
Xは、CH又はNを表す)
を;式(III)の化合物:又はその塩

(式中:
Rは、水素、ハロ、シアノ、C_1-4ハロアルキル、C_1-4ハロアルコキシ、又はSO₂NR⁵R⁶を表し;
R^1a及びR^1bは、独立に、水素又はハロを表し;
R⁵及びR⁶は、独立に、水素又はC_1-4アルキルを表し;かつ
Yは、CH又はNを表す)
と反応させることを含む、方法も提供する。 Accordingly, the present invention provides a process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, comprising a compound of formula (II): or an activated derivative thereof (e.g. acid halide, e.g. acid Chloride or acid anhydride); or its salt

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
X represents CH or N)
A compound of formula (III): or a salt thereof

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl; and
Y represents CH or N)
There is also provided a method comprising reacting with.

(式中:
Rは、水素、ハロ、シアノ、C_1-4ハロアルキル、C_1-4ハロアルコキシ、又はSO₂NR⁵R⁶を表し;
R^1a及びR^1bは、独立に、水素又はハロを表し;
R²は、水素、ハロ、シアノ、C_1-4アルキル、C_1-4アルコキシ、又はC_1-4ハロアルコキシを表し;
R³は、ハロ、シアノ、C_1-4アルキル、又はC_1-4ヒドロキシアルキルを表し;
R⁴は、水素又はC_1-4アルキルを表し;
R⁵及びR⁶は、独立に、水素又はC_1-4アルキルを表し;
Xは、CH又はNを表し;かつ
Yは、CH又はNを表す)
を;式(XI)の化合物:又はその塩

(式中:
Zは、p-トリルSO₂Oなどの脱離基を表す)
と反応させることを含む、方法も提供する。 The present invention relates to a process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, comprising a compound of formula (XV):

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl;
X represents CH or N; and
Y represents CH or N)
A compound of formula (XI): or a salt thereof

(Where:
Z represents a leaving group such as p-tolyl SO ₂ O)
There is also provided a method comprising reacting with.

(式中:
R²は、水素、ハロ、シアノ、C_1-4アルキル、C_1-4アルコキシ、又はC_1-4ハロアルコキシを表し;
R³は、ハロ、シアノ、C_1-4アルキル、又はC_1-4ヒドロキシアルキルを表し;
R⁴は、水素又はC_1-4アルキルを表し;かつ
Xは、CH又はNを表す)
を;式(XXI)の化合物:又はその塩

(式中:
Rは、水素、ハロ、シアノ、C_1-4ハロアルキル、C_1-4ハロアルコキシ、又はSO₂NR⁵R⁶を表し;
R^1a及びR^1bは、独立に、水素又はハロを表し;かつ
R⁵及びR⁶は、独立に、水素又はC_1-4アルキルを表す)
と反応させることを含む、方法も提供する。 The present invention is a process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, comprising a compound of formula (XX): or a salt thereof

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl; and
X represents CH or N)
A compound of formula (XXI): or a salt thereof

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo; and
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl)
There is also provided a method comprising reacting with.

  Pharmaceutically acceptable salts of the compounds of formula (I) include in particular pharmaceutically acceptable acid addition salts of the compounds. Pharmaceutically acceptable acid addition salts of a compound of formula (I) are intended to include therapeutically active non-toxic acid addition salts that the compound of formula (I) can form. These pharmaceutically acceptable acid addition salts can be conveniently obtained by treating the free base form with such a suitable acid in a suitable solvent or solvent mixture. Suitable acids are, for example, inorganic acids such as hydrohalic acids such as hydrochloric acid or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or organic acids such as acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, Pyruvic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid , Including pamonic acid.

  Conversely, the salt form can be converted to the free base form by treatment with a suitable base.

  The definition of a compound of formula (I) is intended to include all tautomers of the compound.

  The definition of a compound of formula (I) is intended to include all solvates of the compound, including solvates of the salts of the compound, unless the context clearly indicates otherwise. Examples of solvates include hydrates.

  The compounds of this disclosure include embodiments in which one or more specified atoms are natural or unnatural isotopes. In one embodiment, the isotope is a stable isotope. Accordingly, the compounds of the present disclosure include, for example, deuterium-containing compounds.

  The present disclosure also extends to all polymorphic forms of the compounds defined herein.

  Novel intermediates described herein, for example compounds of formula (II), (IV), (V), (VII), (VIII), (XIII), (XV), and (XX) and these Salts form a further aspect of the invention. Salts include pharmaceutically acceptable salts (eg, those described above) and pharmaceutically unacceptable salts. Salts of acids (eg, carboxylic acids) include Group 1 and Group 2 metal salts including sodium, potassium, magnesium, and calcium salts.

  In one embodiment, a pharmaceutical composition is provided comprising one or more compounds of the invention, optionally in combination with one or more pharmaceutically acceptable diluents or carriers.

  It is preferred that the compounds of the invention are administered locally to the lung or nose, in particular locally to the lung. Accordingly, in one embodiment, a pharmaceutical composition is provided comprising one or more compounds of the invention, optionally in combination with one or more topically acceptable diluents or carriers.

  Suitable compositions for pulmonary or intranasal administration include powders, liquid solutions, liquid suspensions, nasal drops including solutions or suspensions, or pressurized or non-pressurized aerosols.

  The composition can be conveniently administered in unit dosage form and is described, for example, in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, PA., (1985). Can be prepared by any of the methods well known in the pharmaceutical art. The composition may also be conveniently administered in a multiple unit dosage form.

Topical administration to the nose or lungs can be accomplished by the use of a non-pressurized formulation such as an aqueous solution or suspension. Such formulations can be administered by nebulizers, such as nebulizers that can be handheld and portable, or nebulizers for home or hospital use (ie, non-portable). An example device is the RESPIMAT inhaler. The formulation can include excipients such as water, buffering agents, osmotic pressure adjusting agents, pH adjusting agents, viscosity adjusting agents, surfactants, and co-solvents (eg, ethanol). Suspensions and aerosol formulations (whether pressurized or non-pressurized) usually have a D ₅₀ in finely divided form, for example 0.5-10 μm, for example about 1-5 μm. Contains compounds. The particle size distribution can be expressed using D ₁₀ , D ₅₀ , and D ₉₀ values. The D ₅₀ median of the particle size distribution is defined as the particle size in microns that bisects the distribution. Measurements obtained from laser diffraction are more accurately described as volume distributions, so the D ₅₀ value obtained using this procedure is more meaningfully referred to as the Dv ₅₀ value (median volume distribution). Pass through. As used herein, Dv value refers to the particle size distribution measured using laser diffraction. Similarly, D ₁₀ and D ₉₀ values used in the context of laser diffraction are understood to mean Dv ₁₀ and Dv ₉₀ values, respectively, where 10% of the distribution is located below the D ₁₀ value. , Refers to the particle size where 90% of the distribution is located below the D ₉₀ value.

  According to one particular aspect of the present invention, there is provided a pharmaceutical composition comprising one or more compounds of the present invention in particulate form suspended in an aqueous medium. The aqueous medium usually comprises water and one or more excipients selected from buffers, osmotic pressure adjusting agents, pH adjusting agents, viscosity adjusting agents, and surfactants.

  Topical administration to the nose or lungs can also be accomplished through the use of aerosol formulations. Aerosol formulations usually comprise the active ingredient suspended or dissolved in a suitable aerosol propellant, such as chlorofluorocarbon (CFC) or hydrofluorocarbon (HFC). Suitable CFC propellants include trichloromonofluoromethane (propellant 11), dichlorotetrafluoromethane (propellant 114), and dichlorodifluoromethane (propellant 12). Suitable HFC propellants include tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227). The propellant usually comprises 40% to 99.5% by weight of the total inhalation composition, for example 40% to 90%. The formulation can include an excipient comprising a co-solvent (eg, ethanol) and a surfactant (eg, lecithin, sorbitan trioleate, etc.). Other possible excipients include polyethylene glycol, polyvinyl pyrrolidone, glycerin and the like. The aerosol formulation is packaged in a canister and a suitable dose is delivered by a metered valve (eg, as supplied by Bespak, Valois, or 3M, or by Aptar, Coster, or Vari).

Topical administration to the lung can also be accomplished by use of a dry powder formulation. A dry powder formulation comprises a compound of the present disclosure in finely divided form, usually having an MMD of 1-10 μm or a D ₅₀ of 0.5-10 μm, for example about 1-5 μm. Finely divided forms of powders of the compounds of the invention can be prepared by micronization processes or similar size reduction processes. Micronization can be performed using a jet mill, such as that manufactured by Hosokawa Alpine. The resulting particle size distribution can be measured using laser diffraction (eg, with a Malvern Mastersizer 2000S instrument). The formulation is usually a locally acceptable diluent, such as lactose, glucose, or mannitol, usually a relatively large particle size, eg, 50 μm or more, eg, 100 μm or more MMD or 40-150 μm D ₅₀ (Preferably lactose). As used herein, the term “lactose” refers to α-lactose monohydrate, β-lactose monohydrate, α-lactose anhydride, β-lactose anhydride, and amorphous lactose. Refers to a lactose-containing component. The lactose component can be processed by micronization, sieving, milling, compression, agglomeration, or spray drying. Various forms of commercially available lactose are also included, for example, Lactohale® (inhalation grade lactose; DFE Pharma), InhaLac® 70 (screened lactose for dry powder inhalers; Meggle) , Pharmatose® (DFE Pharma), and Respitose® (sieved inhalation grade lactose; DFE Pharma) products. In one embodiment, the lactose component is selected from the group consisting of α-lactose monohydrate, α-lactose anhydride, and amorphous lactose. Preferably, the lactose is α-lactose monohydrate.

  The dry powder formulation may also contain other excipients such as sodium stearate, calcium stearate, or magnesium stearate.

  Dry powder formulations are typically delivered using a dry powder inhaler (DPI) device. Exemplary dry powder delivery systems include SPINHALER, DISKHALER, TURBOHALER, DISKUS, SKYEHALER, ACCUHALER, and CLICKHALER. Further examples of dry powder delivery systems include ECLIPSE, NEXT, ROTAHALER, HANDIHALER, AEROLISER, CYCLOHALER, BREEZHALER / NEOHALER, MONODOSE, FLOWCAPS, TWINCAPS, X-CAPS, TURBOSPIN, ELPENHALER, MIATHALER, TWISTHALER, NOVOLIZETA, PRESS ORIEL dry powder inhalers, MICRODOSE, PULVINAL, EASYHALER, ULTRAHALER, TAIFUN, PULMOJET, OMNIHALER, GYROHALER, TAPER, CONIX, XCELOVAIR, and PROHALER.

  The compounds of the present invention can be administered topically to another internal or external surface (eg, mucosal surface or skin) or orally. The compounds of the present invention can be conveniently formulated for such administration routes.

  The compounds of the present invention are useful in the treatment of mycosis and for the prevention or treatment of diseases associated with mycosis.

  In one aspect of the invention there is provided the use of one or more compounds of the invention in the manufacture of a medicament for the treatment of mycosis and for the prevention or treatment of diseases associated with mycosis.

  In another aspect of the present invention, there is provided a method of treating a subject having mycosis comprising administering to the subject an effective amount of one or more compounds of the present invention.

  In another aspect of the invention, there is provided a method for the prevention or treatment of a disease associated with mycosis in a subject comprising administering to the subject an effective amount of one or more compounds of the invention. The

  Mycosis can be caused in particular by species of the genus Aspergillus, such as Aspergillus fumigatus or Aspergillus pullulans, in particular Aspergillus fumigatus. Mycosis is a Candida species such as Candida albicans or Candida glabrata, Rhizopus species such as Rhizopus oryzae, Cryptococcus species such as Cryptococcus neoformans, species of the genus Ketomium, e.g., Chaetomium globosum, species of the genus Penicillium, e.g., Penicillium chrysogenum, and species of the genus Trichophyton, e.g. It can also be caused by Trichophyton rubrum.

  A disease associated with mycosis is, for example, pulmonary aspergillosis.

  The compounds of the present invention can be used in prophylactic situations by administering the compound prior to the onset of mycosis.

  Subjects include human and animal subjects, especially human subjects.

  The compounds of the present invention are particularly useful for the treatment of mycosis, eg, Aspergillus fumigatus infection, and for the prevention or treatment of diseases associated with mycosis, eg, Aspergillus fumigatus infection, in subjects at risk. . At-risk subjects include preterm infants, children with congenital defects of the lung or heart, immunocompromised subjects (eg, subjects suffering from HIV infection), asthmatic patients, subjects with cystic fibrosis, elderly subjects And subjects suffering from chronic health disorders affecting the heart or lungs (eg congestive heart failure or chronic obstructive pulmonary disease).

  The compounds of the invention are also useful for the treatment of azole-resistant fungal diseases, such as azole-resistant Aspergillus fumigatus infections, particularly in combination with posaconazole.

  The compounds of the invention can be administered in combination with a second or further active ingredient. Second or further active ingredients are, for example, inhibitors of other antifungal agents (e.g. voriconazole or posaconazole), amphotericin B, echinocandin (e.g. caspofungin), and 3-hydroxy-3-methyl-glutaryl-CoA reductase (Eg, lovastatin, pravastatin, or fluvastatin) can be selected.

  The second or further active ingredient includes a mycosis such as an Aspergillus fumigatus infection or a mycosis such as a disease or mycosis associated with an Aspergillus fumigatus infection such as an Aspergillus fumigatus infection. Active ingredients suitable for the treatment or prevention of are included.

  The compounds of the present invention can be co-formulated with a second or further active ingredient, or the second or further active ingredient can be formulated to be administered separately by the same or different routes .

  For example, the compounds of the present invention can be administered to patients who are already undergoing systemic treatment with antifungal agents such as voriconazole or posaconazole.

  For example, the compound of the present invention comprises one or more selected from amphotericin B, echinocandin, such as caspofungin, and inhibitors of 3-hydroxy-3-methyl-glutaryl-CoA reductase, such as lovastatin, pravastatin, or fluvastatin. It can be co-administered with a drug, eg, co-formulated.

  The compounds of the present invention instead (or additionally) candicidin, philippines, hamycin, natamycin, nystatin, rimocidin, bifonazole, butconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, Oxyconazole, Sertaconazole, Sulconazole, Thioconazole, Albaconazole, Efinaconazole, Epoxyconazole, Fluconazole, Isabconazole, Itraconazole, Propiconazole, Rabconazole, Terconazole, Avafungin, Amorolfine, Butenafin, Naphthyfin, Terfinafin Fungin, Micafungin, benzoic acid, ciclopirox, flucytosine (5- Ruoroshitoshin), griseofulvin, tolnaftate, and one or more agents and co-administration selected from undecylenic acid, for example, can be co-formulated.

  Preferred combination partners include intraconazole, voriconazole, caspofungin, and posaconazole.

  According to one aspect of the present invention, (a) a pharmaceutical composition comprising one or more compounds of the present invention, optionally in combination with one or more diluents or carriers; (b) a second active ingredient, optionally A pharmaceutical composition comprising in combination with one or more diluents or carriers; (c) one or more, optionally each comprising a third or further active ingredient, optionally in combination with one or more diluents or carriers A kit of parts is provided that includes an additional pharmaceutical composition; and (d) instructions for administration of the pharmaceutical composition to a subject in need thereof. A subject in need thereof may or may be susceptible to a mycosis, such as an Aspergillus fumigatus infection.

  The compounds of the invention can be administered at suitable intervals, for example, once a day, twice a day, three times a day, or four times a day.

  Suitable dosages for humans of average body weight (50-70 kg) will be about 50 μg-10 mg / day, for example, 500 μg-5 mg / day, although the exact dose to be administered will be determined by one skilled in the art Can be done.

The compounds of the invention are believed to have one or more of the following advantageous attributes:
-Potent antifungal activity, especially after topical administration to the lungs or nose, in particular against Aspergillus species, such as Aspergillus fumigatus, or Candida species, such as Candida albicans or Candida glabrata, Rhizopus species, such as Rhizopus oryzae, Cryptococcus species, such as Cryptococcus neoformans, Ketmium species, such as Ketmium globosum, Penicillium species, such as Penicillium chrysogenum, or Trichos Activity against species of the phytons, e.g.
Long duration of action in the lung, preferably consistent with once-daily administration;
• low systemic exposure after topical administration to the lung or nose; and • an acceptable safety profile, especially after topical administration to the lung or nose.

(Experimental section)
Abbreviations used herein are defined below (Table 1). Any abbreviation not defined is intended to convey its generally accepted meaning.
Table 1: Abbreviations

(General procedure)
All starting materials and solvents were either obtained from commercial sources or prepared according to literature citations. Unless otherwise stated, all reaction solutions were stirred. The organic solution was routinely dried over anhydrous magnesium sulfate. Hydrogenation was performed on a Thales H-cube flow reactor under the conditions described.

Column chromatography was performed on pre-packed silica (230-400 mesh, 40-63 μm) cartridges using the indicated amounts. SCX was purchased from Supelco. Unless otherwise stated, the reaction mixture to be purified was first diluted with DCM. This solution was loaded directly into SCX and washed with MeOH. The desired material was then eluted by washing with 0.7M NH ₃ in MeOH.

(Preparative reverse phase high performance liquid chromatography)
Method 1: Waters X-Select CSH column C18, 5 μm (19 × 50 mm), flow rate 28 mL min− ¹ , eluted with H ₂ O-MeCN gradient containing 0.1% v / v formic acid over 6.5 min, UV at 254 nm Use detection. Gradient information: 0.0-0.2 min, 20% MeCN; 0.2-5.5 min, increase from 20% MeCN to 80% MeCN; 5.5-5.6 min, increase from 80% MeCN to 95% MeCN; 5.6-6.5 min, 95 Hold at% MeCN.

Method 2: Waters X-Select CSH column C18, 5 μm (19 × 50 mm), flow rate 28 mL min− ¹ , eluted with H ₂ O-MeCN gradient containing 0.1% v / v formic acid over 6.5 min, UV at 254 nm Use detection. Gradient information: 0.0-0.2 minutes, 50% MeCN; 0.2-5.5 minutes, increase from 50% MeCN to 80% MeCN; 5.5-5.6 minutes, increase from 80% MeCN to 95% MeCN; 5.6-6.5 minutes, 95 Hold at% MeCN.

Method 3: Waters X-Select CSH column C18, 5 μm (19 x 50 mm), flow rate 28 mL min- ¹ , eluted with H ₂ O-MeCN gradient containing 0.1% v / v formic acid over 6.5 min, UV at 254 nm Use detection. Gradient information: 0.0-0.2 min, 35% MeCN; 0.2-5.5 min, increase from 35% MeCN to 65% MeCN; 5.5-5.6 min, increase from 65% MeCN to 95% MeCN; 5.6-6.5 min, 95 Hold at% MeCN.

(Analysis method)
(Reverse phase HPLC method :)
Waters Xselect CSH C18 XP column, 2.5 μm (4.6 × 30 mm), 40 ° C .; flow rate 2.5-4.5 mL min- ¹ , 0.1% v / v formic acid (method a) or 10 mM NH ₄ HCO _{3 in} water (method b) Elute with either H ₂ O-MeCN gradient over 4 min and utilize UV detection at 254 nm. Gradient information: 0 to 3.00 min, increase from 95% H ₂ O-5% MeCN to 5% H ₂ O-95% MeCN; hold at 3.00 to 3.01 min, 5% H ₂ O-95% MeCN, flow rate the increase in 4.5mL min ^-1; 3.01 to 3.50 minutes, hold at _{5% H 2 O-95%} MeCN; 3.50~3.60 minutes, return to _{95% H 2 O-5%} MeCN, 3.50mL min flow rate reduced to ^-1; 3.60 to 3.90 minutes, hold at _{95% H 2 O-5%} MeCN; 3.90~4.00 minutes, held at _{95% H 2 O-5%} MeCN, flow rate to 2.5mL min ^-1 Decrease.

( ¹ H NMR spectroscopy :)
¹ H NMR spectra were acquired on a Bruker Advance III spectrometer at 400 MHz using the remaining non-deuterated solvent as a reference and run in DMSO-d ₆ unless otherwise specified.

tert-ブチルピペラジン-1-カルボキシレート(7.44g、40.0mmol)、4-ブロモ-2-メチルフェノール(Xa)(6.23g、33.3mmol)、RuPhos(311mg、0.67mmol)、及びRuPhos G3(557mg、0.67mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した。LiHMDSの溶液(THF中1M、100mL、100mmol)を添加し、反応混合物を70℃で3時間加熱した。RTに冷却した後、該混合物を1M塩酸(100mL)の添加によりクエンチし、その後、1M水性NaHCO₃(100mL)で中性化した。水層をEtOAc(3×100mL)で抽出し、合わせた有機抽出物を乾燥させた。揮発性物質を真空中で除去すると、粗生成物が得られ、これを、フラッシュカラムクロマトグラフィー(SiO₂、120g、イソヘキサン中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(XIIa)が薄茶色の固体(7.80g、78％)として得られた; R^t 2.07分(方法b); m/z 293(M+H)⁺(ES⁺);

(Representative procedure for the preparation of intermediates)
(tert-butyl 4- (4-hydroxy-3-methylphenyl) piperazine-1-carboxylate)

tert-butyl piperazine-1-carboxylate (7.44 g, 40.0 mmol), 4-bromo-2-methylphenol (Xa) (6.23 g, 33.3 mmol), RuPhos (311 mg, 0.67 mmol), and RuPhos G3 (557 mg, 0.67 mmol) was evacuated from the flask and refilled with nitrogen three times. A solution of LiHMDS (1M in THF, 100 mL, 100 mmol) was added and the reaction mixture was heated at 70 ° C. for 3 h. After cooling to RT, the mixture was quenched by the addition of 1M hydrochloric acid (100 mL) and then neutralized with 1M aqueous NaHCO ₃ (100 mL). The aqueous layer was extracted with EtOAc (3 × 100 mL) and the combined organic extracts were dried. If the volatiles are removed in vacuo, the crude product is obtained which, by flash column chromatography _{(SiO 2, 120g, 0~100%} EtOAc in isohexane, gradient elution) to give the intermediate title compound (XIIa) was obtained as a light brown solid (7.80 g, 78%); R ^t 2.07 min (method b); m / z 293 (M + H) ⁺ (ES ⁺ );

中間体(XIIa)(7.80g、25.1mmol)のDMSO(60mL)溶液に、水性水酸化ナトリウム(3.0mL、12.5M、37.6mmol)を添加した。混合物をRTで10分間撹拌し、その後、トシレート(XIa)(APIChem製、カタログ番号: AC-8330、12.4g、27.6mmol)で少しずつ処理した。反応混合物を30℃で18時間撹拌し、RTに冷却し、水(200mL)を添加した。得られた混合物をEtOAc(3×200mL)で抽出し、合わせた有機抽出物をブライン(2×200mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、茶色の油状物が得られた。¹H NMRによる粗Boc保護生成物(VIIa)の分析により、該生成物が排出副生成物として形成された、〜10％のアルケン:(R)-1-((2-(2,4-ジフルオロフェニル)-4-メチレンテトラヒドロフラン-2-イル)メチル)-1H-1,2,4-トリアゾールを含むことが示された。粗ウレタン(VIIa)をDCM(150mL)中に取り、TFA(39.0mL、502mmol)で処理した。RTで2時間後、反応混合物を真空中で濃縮して、揮発性物質の大部分を除去し、その後、EtOAc(200mL)で希釈し、水性NaOH(2M、200mL)で洗浄した。水相を分離し、EtOAc(2×200mL)で抽出した。合わせた有機抽出物をブライン(2×200mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、薄茶色の油状物が得られた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、80g、DCM中0〜10％の0.7M NH₃/MeOH、勾配溶出)により精製すると、表題化合物の中間体(Va)が粘性のある薄茶色の油状物(9.46g、80％)として得られた; R^t 1.91分(方法b); m/z 470(M+H)⁺(ES⁺);

(1- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3-yl ) Methoxy) -3-methylphenyl) piperazine

To a solution of intermediate (XIIa) (7.80 g, 25.1 mmol) in DMSO (60 mL) was added aqueous sodium hydroxide (3.0 mL, 12.5 M, 37.6 mmol). The mixture was stirred at RT for 10 min and then treated in portions with tosylate (XIa) (APIChem, catalog number: AC-8330, 12.4 g, 27.6 mmol). The reaction mixture was stirred at 30 ° C. for 18 hours, cooled to RT and water (200 mL) was added. The resulting mixture is extracted with EtOAc (3 × 200 mL) and the combined organic extracts are washed with brine (2 × 200 mL), then dried and evaporated in vacuo to give a brown oil. It was. Analysis of the crude Boc protected product (VIIa) by ¹ H NMR revealed that the product was formed as an effluent by-product of ˜10% alkene: (R) -1-((2- (2,4- Difluorophenyl) -4-methylenetetrahydrofuran-2-yl) methyl) -1H-1,2,4-triazole was shown to contain. The crude urethane (VIIa) was taken up in DCM (150 mL) and treated with TFA (39.0 mL, 502 mmol). After 2 h at RT, the reaction mixture was concentrated in vacuo to remove most of the volatiles, then diluted with EtOAc (200 mL) and washed with aqueous NaOH (2M, 200 mL). The aqueous phase was separated and extracted with EtOAc (2 × 200 mL). The combined organic extracts were washed with brine (2 × 200 mL), then dried and evaporated in vacuo to give a light brown oil. The crude product was purified by flash column chromatography _{(SiO 2, 80g, 0.7M NH} 3 / MeOH 0-10% in DCM, gradient elution) to give the intermediate title compound (Va) of pale brown viscous Obtained as an oil (9.46 g, 80%); R ^t 1.91 min (method b); m / z 470 (M + H) ⁺ (ES ⁺ );

中間体(Va)(9.00g、19.2mmol)、メチル-4-ブロモベンゾエート(VIa)(4.95g、23.0mmol)、RuPhos(0.18g、0.38mmol、2mol％)、RuPhosG3(0.32g、0.38mmol、2mol％)、及び炭酸セシウム(9.99g、30.7mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した後、DMF(150mL)を添加した。混合物を80℃で22時間加熱し、その後、まだ熱いうちに、水(150mL)に注ぎ入れると、茶色のゴム状物質が形成した。さらなる水(300mL)を添加し、水相をDCM(2×200mL)で抽出しした。有機抽出物を合わせ、真空中で濃縮すると、茶色の油状物が得られ、これを水(100mL)に注ぎ入れた。得られた沈殿物を濾過により回収し、その後、THF(100mL)に再懸濁させた。混合物を還流状態で1時間加熱し、その間に、クリーム色の懸濁液が形成された。混合物をRTに冷却し、得られた沈殿物を濾過により回収し、THF(2×50mL)で洗浄し、その後、真空中で乾燥させると、表題化合物の中間体(IVa)が薄黄色の固体(9.48g、79％)として得られた; R^t 2.79分(方法b); m/z 604(M+H)⁺(ES⁺);

(Methyl 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran -3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) benzoate)

Intermediate (Va) (9.00 g, 19.2 mmol), methyl-4-bromobenzoate (VIa) (4.95 g, 23.0 mmol), RuPhos (0.18 g, 0.38 mmol, 2 mol%), RuPhosG3 (0.32 g, 0.38 mmol, 2 mol%) and cesium carbonate (9.99 g, 30.7 mmol) were evacuated from the flask and refilled with nitrogen three times before adding DMF (150 mL). The mixture was heated at 80 ° C. for 22 hours and then poured into water (150 mL) while still hot to form a brown gum. Additional water (300 mL) was added and the aqueous phase was extracted with DCM (2 × 200 mL). The organic extracts were combined and concentrated in vacuo to give a brown oil that was poured into water (100 mL). The resulting precipitate was collected by filtration and then resuspended in THF (100 mL). The mixture was heated at reflux for 1 hour, during which time a creamy suspension formed. The mixture was cooled to RT and the resulting precipitate was collected by filtration, washed with THF (2 × 50 mL) and then dried in vacuo to yield the title compound intermediate (IVa) as a pale yellow solid. (9.48 g, 79%); R ^t 2.79 min (method b); m / z 604 (M + H) ⁺ (ES ⁺ );

エチル 4-(ピペラジン-1-イル)ベンゾエート(XIVa)(20.0g、85.0mmol)及び4-ブロモ-2-メチルフェノール(Xa)(19.2g、102mmol)のDMF(213mL)溶液を仕込んだフラスコから空気を排出し、窒素を3回再充填した。RuPhos G3(1.43g、1.71mmol)を添加し、フラスコから空気を排出し、窒素を再充填した。反応混合物を0℃に冷却し、LiHMDS(17.1g、102mmol)を添加した。反応液をRTで10分間撹拌し、その後、水浴中で冷却し、LiHMDS(20.0g、120mmol)を同じ分量(7×2.85g)で5分間の間隔で添加した。得られた溶液をRTで30分間撹拌し、その後、0℃に冷却し、2M塩酸(200mL)で処理して、6〜7のpHを生じさせた。混合物をRTで15分間撹拌し、その後、EtOAc(220mL)で抽出した。水層を分離し、EtOAc(4×50mL)で抽出し、合わせた有機物をブライン(6×50mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、クリーム色の固体が得られた。イソヘキサンとIPA(1:1、150mL)の混合物を添加し、懸濁液をRTで30分間撹拌した。固体を濾過により回収し、フィルターケーキをイソヘキサンとIPA(1:1、2×10mL)の混合物、次いで、イソヘキサン(4×10mL)で洗浄し、真空中、40℃で18時間で乾燥させると、表題化合物の中間体(XIIIa)がクリーム色の固体(15.3g、50％)として得られた; R^t 2.29分(方法b); m/z 341(M+H)⁺(ES⁺);

(Ethyl 4- (4- (4-hydroxy-3-methylphenyl) piperazin-1-yl) benzoate)

From a flask charged with a solution of ethyl 4- (piperazin-1-yl) benzoate (XIVa) (20.0 g, 85.0 mmol) and 4-bromo-2-methylphenol (Xa) (19.2 g, 102 mmol) in DMF (213 mL) The air was evacuated and refilled with nitrogen three times. RuPhos G3 (1.43 g, 1.71 mmol) was added and the flask was evacuated and refilled with nitrogen. The reaction mixture was cooled to 0 ° C. and LiHMDS (17.1 g, 102 mmol) was added. The reaction was stirred at RT for 10 min, then cooled in a water bath and LiHMDS (20.0 g, 120 mmol) was added in equal portions (7 × 2.85 g) at 5 min intervals. The resulting solution was stirred at RT for 30 minutes, then cooled to 0 ° C. and treated with 2M hydrochloric acid (200 mL) to give a pH of 6-7. The mixture was stirred at RT for 15 minutes and then extracted with EtOAc (220 mL). The aqueous layer was separated and extracted with EtOAc (4 × 50 mL) and the combined organics were washed with brine (6 × 50 mL) then dried and evaporated in vacuo to give a cream colored solid . A mixture of isohexane and IPA (1: 1, 150 mL) was added and the suspension was stirred at RT for 30 min. The solid was collected by filtration and the filter cake was washed with a mixture of isohexane and IPA (1: 1, 2 × 10 mL), then isohexane (4 × 10 mL), and dried in vacuo at 40 ° C. for 18 hours. The title compound intermediate (XIIIa) was obtained as a cream colored solid (15.3 g, 50%); R ^t 2.29 min (method b); m / z 341 (M + H) ⁺ (ES ⁺ );

0℃に冷却した中間体(XIIIa)(15.3g、44.9mmol)のDMF(110mL)溶液に、ナトリウムエトキシド(3.13g、46.1mmol)を添加し、混合物を0℃で10分間撹拌し、その後、トシレート(XIa)で処理した。反応混合物をRTに温めておき、50℃に1時間加熱し、その後、RTに冷却した。塩酸(1M、60mL)及び水(200mL)を添加し、混合物をRTで30分間撹拌し、その後、DCM(150mL)で抽出した。水層を分離し、DCM(2×50mL)で抽出し、合わせた有機物をブライン(4×30mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、クリーム色の固体が得られた。この固体をイソヘキサンとIPA(80mL)の等量混合物に懸濁させ、RTで1時間撹拌した。固体を濾過により回収し、イソヘキサンとIPA(3×20mL)の1:1の混合物で洗浄し、その後、真空中、40℃で18時間乾燥させると、表題化合物の中間体(IVb)が白色の固体(16.4g、56％)として得られた; R^t 2.92分(方法b); m/z 618(M+H)⁺(ES⁺);

(Ethyl 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran -3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) benzoate)

To a solution of intermediate (XIIIa) (15.3 g, 44.9 mmol) in DMF (110 mL) cooled to 0 ° C., sodium ethoxide (3.13 g, 46.1 mmol) is added and the mixture is stirred at 0 ° C. for 10 minutes, after which And treated with tosylate (XIa). The reaction mixture was allowed to warm to RT and heated to 50 ° C. for 1 hour and then cooled to RT. Hydrochloric acid (1M, 60 mL) and water (200 mL) were added and the mixture was stirred at RT for 30 min and then extracted with DCM (150 mL). The aqueous layer was separated and extracted with DCM (2 × 50 mL) and the combined organics were washed with brine (4 × 30 mL) then dried and evaporated in vacuo to give a cream colored solid . This solid was suspended in an equal mixture of isohexane and IPA (80 mL) and stirred at RT for 1 h. The solid was collected by filtration, washed with a 1: 1 mixture of isohexane and IPA (3 × 20 mL), and then dried in vacuo at 40 ° C. for 18 hours to yield the title compound intermediate (IVb) as a white Obtained as a solid (16.4 g, 56%); R ^t 2.92 min (method b); m / z 618 (M + H) ⁺ (ES ⁺ );

5-ブロモ-2-ヒドロキシベンゾニトリル(Xb)(1.00g、5.05mmol)、臭化テトラブチルアンモニウム(814mg、2.53mmol)、及びリン酸カリウム三塩基一水和物(1.16g、5.05mmol)の撹拌した水(10mL)懸濁液に、臭化ベンジル(600μL、5.05mmol)を添加した。混合物をRTで18時間撹拌し、DCM(3×20mL)で抽出した。合わせた有機物をブライン(20mL)で洗浄し、その後、乾燥させ、真空中で蒸発させた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、イソヘキサン中0〜40％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(Xc)が白色の固体(1.22g、82％)として得られた; R^t 2.53分(方法a); m/z、イオン化は認められなかった;

(2- (Benzyloxy) -5-bromobenzonitrile)

Of 5-bromo-2-hydroxybenzonitrile (Xb) (1.00 g, 5.05 mmol), tetrabutylammonium bromide (814 mg, 2.53 mmol), and potassium phosphate tribasic monohydrate (1.16 g, 5.05 mmol). To a stirred suspension of water (10 mL) was added benzyl bromide (600 μL, 5.05 mmol). The mixture was stirred at RT for 18 hours and extracted with DCM (3 × 20 mL). The combined organics were washed with brine (20 mL) then dried and evaporated in vacuo. So obtained crude product was purified by flash column chromatography _{(SiO 2, 12g, 0~40%} EtOAc in isohexane, gradient elution) to give the intermediate title compound (Xc) as a white solid (1.22 g, 82%); R ^t 2.53 min (method a); m / z, no ionization observed;

メチル 4-(ピペラジン-1-イル)ベンゾエート(XIVa)(459mg、2.08mmol)、中間体(Xc)(500mg、1.74mmol)、RuPhos(40.0mg、87.0μmol)、RuPhos G3(67.0mg、87.0μmol)、及び炭酸セシウム(678mg、2.08mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した後、DMF(8.0mL)を添加した。混合物を80℃で18時間加熱し、その後、RTに冷却し、水(50mL)とEtOAc(50mL)に分配した。有機相を分離し、保持し、水層をEtOAc(2×50mL)で抽出した。合わせた有機物をブライン(3×50mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、黄色の固体が得られた。粗生成物をジエチルエーテル(20mL)中で粉砕し、濾過により回収し、真空中、40℃で18時間乾燥させると、表題化合物(XIIIb)がベージュ色の固体(286mg、37％)として得られた; R^t 2.74分(方法a); m/z 428(M+H)⁺(ES⁺);

(Methyl 4- (4- (4- (benzyloxy) -3-cyanophenyl) piperazin-1-yl) benzoate)

Methyl 4- (piperazin-1-yl) benzoate (XIVa) (459 mg, 2.08 mmol), intermediate (Xc) (500 mg, 1.74 mmol), RuPhos (40.0 mg, 87.0 μmol), RuPhos G3 (67.0 mg, 87.0 μmol) ) And cesium carbonate (678 mg, 2.08 mmol) were evacuated and refilled with nitrogen three times before adding DMF (8.0 mL). The mixture was heated at 80 ° C. for 18 hours, then cooled to RT and partitioned between water (50 mL) and EtOAc (50 mL). The organic phase was separated and retained and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organics were washed with brine (3 × 50 mL), then dried and evaporated in vacuo to give a yellow solid. The crude product was triturated in diethyl ether (20 mL), collected by filtration and dried in vacuo at 40 ° C. for 18 hours to give the title compound (XIIIb) as a beige solid (286 mg, 37%). R ^t 2.74 min (method a); m / z 428 (M + H) ⁺ (ES ⁺ );

中間体(XIIIb)(286mg、0.669mmol)のEtOAc(80mL)溶液を、H-Cube(10％パラジウム炭、70×4mm、完全水素、30℃、1mL/分)を用いて水素化した。溶媒を真空中で蒸発させ、残渣をフラッシュカラムクロマトグラフィー(SiO₂、4g、DCM中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(XIIIc)が白色の固体(139mg、60％)として得られた; R^t 2.06分(方法a); m/z 338(M+H)⁺(ES⁺);

(Methyl 4- (4- (3-cyano-4-hydroxyphenyl) piperazin-1-yl) benzoate)

A solution of intermediate (XIIIb) (286 mg, 0.669 mmol) in EtOAc (80 mL) was hydrogenated using H-Cube (10% palladium on charcoal, 70 × 4 mm, complete hydrogen, 30 ° C., 1 mL / min). The solvent was evaporated in vacuo, the residue was purified by flash column chromatography _{(SiO 2, 4g, 0~100%} EtOAc in DCM, gradient elution) to give the intermediate title compound (XIIIc) as a white solid (139 mg, 60%); R ^t 2.06 min (method a); m / z 338 (M + H) ⁺ (ES ⁺ );

中間体(XIIIc)(139mg、0.412mmol)のDMSO(1.0mL)溶液に、水性水酸化ナトリウム(30.0μL、12.5M、0.381mmol)を添加し、混合物をRTで10分間撹拌し、その後、トシレート(XIa)(154mg、0.343mmol)のDMSO(1.0mL)溶液で処理した。反応混合物を30℃で18時間撹拌し、RTに冷却し、水(30mL)を添加した。得られた混合物をEtOAc(3×50mL)で抽出し、合わせた有機抽出物をブライン(2×30mL)で洗浄し、乾燥させ、真空中で蒸発させると、茶色の油状物が得られた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、DCM中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(IVc)が白色の泡状物質(100mg、46％)として得られた; R^t 2.59分(方法a); m/z 615(M+H)⁺(ES⁺);

(Methyl 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran -3-yl) methoxy) -3-cyanophenyl) piperazin-1-yl) benzoate)

To a solution of intermediate (XIIIc) (139 mg, 0.412 mmol) in DMSO (1.0 mL) was added aqueous sodium hydroxide (30.0 μL, 12.5 M, 0.381 mmol) and the mixture was stirred at RT for 10 min, then tosylate. Treated with a solution of (XIa) (154 mg, 0.343 mmol) in DMSO (1.0 mL). The reaction mixture was stirred at 30 ° C. for 18 hours, cooled to RT and water (30 mL) was added. The resulting mixture was extracted with EtOAc (3 × 50 mL) and the combined organic extracts were washed with brine (2 × 30 mL), dried and evaporated in vacuo to give a brown oil. The crude product was purified by flash column chromatography _{(SiO 2, 12g, 0~100%} EtOAc in DCM, gradient elution) to give, as an intermediate for the title compound (IVc) as a white foam (100 mg, 46%) Obtained; R ^t 2.59 min (method a); m / z 615 (M + H) ⁺ (ES ⁺ );

4-ブロモ-2-メチルフェノール(Xa)(920mg、4.89mmol)のDMSO(10mL)溶液に、水性水酸化ナトリウム(0.39mL、12.5M、4.89mmol)を添加し、混合物をRTで10分間撹拌し、その後、トシレート(XIa)(2.00g、4.45mmol)で処理した。反応混合物を60℃で72時間撹拌し、その後、RTに冷却し、水(25mL)とEtOAc(20mL)に分配した。有機相を分離し、保持し、水層をEtOAc(3×25mL)で抽出した。合わせた有機抽出物をブライン(3×15mL)で洗浄し、その後、乾燥させ、真空中で蒸発させた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、DCM中0〜30％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(VIIIa)が無色の油状物(1.84g、86％)として得られた; R^t 2.78分(方法a); m/z 464(M+H)⁺(ES⁺);

(1-(((2R, 4R) -4-((4-Bromo-2-methylphenoxy) methyl) -2- (2,4-difluorophenyl) tetrahydrofuran-2-yl) methyl) -1H-1, 2,4-triazole)

To a solution of 4-bromo-2-methylphenol (Xa) (920 mg, 4.89 mmol) in DMSO (10 mL) is added aqueous sodium hydroxide (0.39 mL, 12.5 M, 4.89 mmol) and the mixture is stirred at RT for 10 min. And then treated with tosylate (XIa) (2.00 g, 4.45 mmol). The reaction mixture was stirred at 60 ° C. for 72 hours, then cooled to RT and partitioned between water (25 mL) and EtOAc (20 mL). The organic phase was separated and retained and the aqueous layer was extracted with EtOAc (3 × 25 mL). The combined organic extracts were washed with brine (3 × 15 mL) then dried and evaporated in vacuo. The crude product was purified by flash column chromatography _{(SiO 2, 12g, 0~30%} EtOAc in DCM, gradient elution) to give the intermediate title compound (VIIIa) is a colorless oil (1.84 g, 86%) as a Obtained; R ^t 2.78 min (method a); m / z 464 (M + H) ⁺ (ES ⁺ );

エチル 4-(ピペラジン-1-イル)ベンゾエート(XIVa)(103mg、0.44mmol)、中間体(VIIIa)(170mg、0.37mmol)、RuPhos(8.5mg、18μmol)、RuPhos G3(14.2mg、18μmol)、及び炭酸セシウム(191mg、0.59mmol)を仕込んだバイアルから空気を排出し、窒素を3回再充填した後、DMF(3.0mL)を添加した。混合物を80℃で18時間、その後、100℃で24時間加熱した。反応混合物をRTに冷却し、水(10mL)とEtOAc(10mL)に分配した。有機相を分離し、保持し、水層をEtOAc(3×10mL)で抽出した。合わせた有機物をブライン(3×10mL)で洗浄し、その後、乾燥させ、真空中で蒸発させた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、イソヘキサン中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(IVb)が白色の固体(100mg、43％)として得られた。 (Ethyl 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran -3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) benzoate)

Ethyl 4- (piperazin-1-yl) benzoate (XIVa) (103 mg, 0.44 mmol), intermediate (VIIIa) (170 mg, 0.37 mmol), RuPhos (8.5 mg, 18 μmol), RuPhos G3 (14.2 mg, 18 μmol), And air was evacuated from the vial charged with cesium carbonate (191 mg, 0.59 mmol) and refilled with nitrogen three times before adding DMF (3.0 mL). The mixture was heated at 80 ° C. for 18 hours and then at 100 ° C. for 24 hours. The reaction mixture was cooled to RT and partitioned between water (10 mL) and EtOAc (10 mL). The organic phase was separated and retained and the aqueous layer was extracted with EtOAc (3 × 10 mL). The combined organics were washed with brine (3 × 10 mL) then dried and evaporated in vacuo. The crude product was purified by flash column chromatography (SiO _2, 12 g, isohexane 0 to 100% EtOAc, gradient elution) to give the intermediate title compound (IVb) is obtained as a white solid (100 mg, 43%) It was.

6-ブロモ-2-メチルピリジン-3-オール(Xd)(1.00g、5.32mmol)のDMSO(17mL)溶液に、水性水酸化ナトリウム(2.80mL、2.0M、5.32mmol)を添加し、混合物をRTで10分間撹拌し、その後、トシレート(XIa)(2.17g、4.84mmol)で少しずつ処理した。反応混合物を65℃で18時間撹拌し、RTに冷却し、水(30mL)を添加した。得られた混合物をEtOAc(3×50mL)で抽出し、合わせた有機物をブライン(2×30mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、黄色の油状物が得られた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、40g、DCM中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(VIIIb)が白色の固体(1.50g、64％)として得られた; R^t 2.28分(方法a); m/z 465/467(M+H)⁺(ES⁺);

(3-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3-yl) methoxy) -6-Bromo-2-methylpyridine)

To a solution of 6-bromo-2-methylpyridin-3-ol (Xd) (1.00 g, 5.32 mmol) in DMSO (17 mL) was added aqueous sodium hydroxide (2.80 mL, 2.0 M, 5.32 mmol) and the mixture was Stir at RT for 10 min, then treat with tosylate (XIa) (2.17 g, 4.84 mmol) in portions. The reaction mixture was stirred at 65 ° C. for 18 hours, cooled to RT and water (30 mL) was added. The resulting mixture was extracted with EtOAc (3 × 50 mL) and the combined organics were washed with brine (2 × 30 mL), then dried and evaporated in vacuo to give a yellow oil. The crude product was purified by flash column chromatography _{(SiO 2, 40g, 0~100%} EtOAc in DCM, gradient elution) to afford to give the title intermediate compound (VIIIb) as a white solid (1.50 g, 64%) as a It was; R ^t 2.28 min (method a); m / z 465/467 ( M + H) + (ES +);

tert-ブチルピペラジン-1-カルボキシレート(IXa)(550mg、2.93mmol)、中間体(VIIIb)(1.50g、3.22mmol)、RuPhos(68.0mg、0.147mmol)、RuPhos G3(123mg、0.147mmol)、及び炭酸セシウム(1.53g、4.69mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した後、DMF(15mL)を添加した。混合物を80℃で18時間加熱し、その後、RTに冷却し、水(100mL)とEtOAc(100mL)に分配した。有機相を分離し、保持し、水層をEtOAc(2×100mL)で抽出した。合わせた有機抽出物をブライン(3×50mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、黄色の油状物が得られた。粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、80g、DCM中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(VIIb)が白色の泡状物質(1.38g、77％)として得られた; R^t 2.14分(方法a); m/z 571(M+H)⁺(ES⁺);

(tert-Butyl 4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran- 3-yl) methoxy) -6-methylpyridin-2-yl) piperazine-1-carboxylate)

tert-butylpiperazine-1-carboxylate (IXa) (550 mg, 2.93 mmol), intermediate (VIIIb) (1.50 g, 3.22 mmol), RuPhos (68.0 mg, 0.147 mmol), RuPhos G3 (123 mg, 0.147 mmol), And evacuated from the flask charged with cesium carbonate (1.53 g, 4.69 mmol) and refilled with nitrogen three times before adding DMF (15 mL). The mixture was heated at 80 ° C. for 18 hours, then cooled to RT and partitioned between water (100 mL) and EtOAc (100 mL). The organic phase was separated and retained and the aqueous layer was extracted with EtOAc (2 × 100 mL). The combined organic extracts were washed with brine (3 × 50 mL) then dried and evaporated in vacuo to give a yellow oil. The crude product was purified by flash column chromatography _{(SiO 2, 80g, 0~100%} EtOAc in DCM, gradient elution) to give the intermediate title compound (VIIb) as a white foam (1.38 g, 77%) obtained as; R ^t 2.14 min (method a); m / z 571 ( M + H) + (ES +);

中間体(VIIb)のDCM(10mL)溶液をTFA(2.62mL、34.0mmol)で処理し、RTで2時間撹拌した。反応混合物を真空中で濃縮すると、茶色の油状物が得られ、これをEtOAc(50mL)と1M水性NaHCO₃(20mL)に分配した。有機層を分離し、ブライン(20mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、表題化合物の中間体(Vb)が茶色の泡状物質(858mg、76％)として得られた; R^t 1.26分(方法a); m/z 471(M+H)⁺(ES⁺);

(1- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3-yl ) Methoxy) -6-methylpyridin-2-yl) piperazine

A solution of intermediate (VIIb) in DCM (10 mL) was treated with TFA (2.62 mL, 34.0 mmol) and stirred at RT for 2 h. The reaction mixture was concentrated in vacuo to give a brown oil which was partitioned between EtOAc (50 mL) and 1M aqueous NaHCO ₃ (20 mL). The organic layer was separated, washed with brine (20 mL), then dried and evaporated in vacuo to give the title compound intermediate (Vb) as a brown foam (858 mg, 76%). R ^t 1.26 min (method a); m / z 471 (M + H) ⁺ (ES ⁺ );

中間体(Vb)(860mg、1.82mmol)、メチル-4-ブロモベンゾエート(VIa)(470mg、2.19mmol)、RuPhos(17.0mg、0.0360mmol、2mol％)、RuPhosG3(28.0mg、0.0360mmol、2mol％)、及び炭酸セシウム(950mg、2.92mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した後、DMF(6.0mL)を添加した。混合物を80℃で18時間加熱し、その後、RTに冷却し、EtOAc(50mL)と水(50mL)に分配した。有機層を分離し、ブライン(3×50mL)で洗浄し、その後、乾燥させ、真空中で蒸発させると、ベージュ色の固体が得られた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、24g、DCM中0〜100％EtOAc、勾配溶出)により精製すると、表題化合物の中間体(IVd)が黄色の固体(690mg、61％)として得られた; R^t 2.21分(方法a); m/z 605(M+H)⁺(ES⁺);

(Methyl 4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran -3-yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) benzoate)

Intermediate (Vb) (860 mg, 1.82 mmol), methyl-4-bromobenzoate (VIa) (470 mg, 2.19 mmol), RuPhos (17.0 mg, 0.0360 mmol, 2 mol%), RuPhosG3 (28.0 mg, 0.0360 mmol, 2 mol%) ) And cesium carbonate (950 mg, 2.92 mmol) were evacuated and refilled with nitrogen three times before adding DMF (6.0 mL). The mixture was heated at 80 ° C. for 18 hours, then cooled to RT and partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was separated and washed with brine (3 × 50 mL), then dried and evaporated in vacuo to give a beige solid. The crude product so obtained was purified by flash column chromatography (SiO ₂ , 24 g, 0-100% EtOAc in DCM, gradient elution) to yield the title compound intermediate (IVd) as a yellow solid (690 mg , 61%); R ^t 2.21 min (method a); m / z 605 (M + H) ⁺ (ES ⁺ );

中間体(IVa)(9.00g、14.9mmol)のDMSO(370mL)懸濁液に、水酸化リチウム(1.79g、74.5mmol)の水(37.0mL)溶液を添加した。混合物を70℃で22時間加熱し、その後、RTに冷却し、水(1000mL)で希釈し、1M水性塩酸(80mL)の添加により、(〜pH 2に)酸性化した。混合物を氷浴中で2時間冷却し、得られた沈殿物を濾過により回収した。フィルターケーキを水(3×80mL)で洗浄し、真空中、50℃で乾燥させると、表題化合物の中間体(IIa)が白色の固体(4.66g、54％)として得られた; R^t 2.21分(方法1a); m/z 590(M+H)⁺(ES⁺);

(4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran- 3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) benzoic acid)
(Hydrolysis of methyl ester (IVa))

To a suspension of intermediate (IVa) (9.00 g, 14.9 mmol) in DMSO (370 mL) was added a solution of lithium hydroxide (1.79 g, 74.5 mmol) in water (37.0 mL). The mixture was heated at 70 ° C. for 22 hours, then cooled to RT, diluted with water (1000 mL) and acidified (to pH 2) by addition of 1M aqueous hydrochloric acid (80 mL). The mixture was cooled in an ice bath for 2 hours and the resulting precipitate was collected by filtration. The filter cake was washed with water (3 × 80 mL) and dried in vacuo at 50 ° C. to give the title compound intermediate (IIa) as a white solid (4.66 g, 54%); R ^t 2.21 Minutes (Method 1a); m / z 590 (M + H) ⁺ (ES ⁺ );

(Hydrolysis of ethyl ester (IVb))
To a suspension of intermediate (IVb) (16.4 g, 26.6 mmol) in DMSO (375 mL) was added a solution of lithium hydroxide (3.18 g, 74.5 mmol) in water (50 mL). The mixture was heated at 70 ° C. for 22 hours, then cooled to RT, poured into water (500 mL) and acidified (to pH 5-6) by addition of 2M hydrochloric acid (70 mL). The mixture was stirred at RT for 30 min and the resulting solid was collected by filtration, washed with water (2 × 20 mL) and diethyl ether (3 × 30 mL) and then dried in vacuo at 40 ° C. for 18 hours. The intermediate (IIa) of the title compound was obtained as a brown solid (14.2 g, 84%); R ^t 2.26 min (Method 1a); m / z 590 (M + H) ⁺ (ES ⁺ );

中間体(IVc)(100mg、0.163mmol)のDMSO(8.0mL)懸濁液に、水酸化リチウム(19mg、0.81mmol)の水(1.0mL)溶液を添加した。混合物を50℃で18時間加熱し、その後、RTに冷却し、水(10mL)で希釈し、1M塩酸(2.0mL)の添加により、(〜pH 3に)酸性化した。混合物を4:1のDCM/EtOAc(2×25mL)で抽出し、合わせた有機物を水(2×10mL)で洗浄し、乾燥させ、真空中で蒸発させると、白色の固体が得られ、これを、真空中、40℃で乾燥させると、表題化合物の中間体(IIb)が白色の固体(74mg、75％)として得られた; R^t 2.32分(方法a); m/z 601(M+H)⁺(ES⁺);

(4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran- 3-yl) methoxy) -3-cyanophenyl) piperazin-1-yl) benzoic acid)

To a suspension of intermediate (IVc) (100 mg, 0.163 mmol) in DMSO (8.0 mL) was added a solution of lithium hydroxide (19 mg, 0.81 mmol) in water (1.0 mL). The mixture was heated at 50 ° C. for 18 hours then cooled to RT, diluted with water (10 mL) and acidified (to pH 3) by addition of 1M hydrochloric acid (2.0 mL). The mixture was extracted with 4: 1 DCM / EtOAc (2 × 25 mL) and the combined organics were washed with water (2 × 10 mL), dried and evaporated in vacuo to give a white solid which Was dried in vacuo at 40 ° C. to give the title compound intermediate (IIb) as a white solid (74 mg, 75%); R ^t 2.32 min (method a); m / z 601 (M + H) ⁺ (ES ⁺ );

中間体(IVd)(690mg、1.14mmol)のDMSO(54mL)懸濁液に、水酸化リチウム(140mg、5.71mmol)の水(9.0mL)溶液を添加した。混合物を70℃で22時間加熱し、その後、RTに冷却し、水(100mL)で希釈し、1M塩酸(6.0mL)の添加により、(〜pH 2に)酸性化した。混合物を氷浴中で15分間冷却し、得られた沈殿物を濾過により回収し、水(3×50mL)で洗浄し、真空中、40℃で乾燥させると、表題化合物の中間体(IIc)が黄色の固体(617mg、87％)として得られた; R^t 1.91分(方法a); m/z 591(M+H)⁺(ES⁺);

(4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran- 3-yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) benzoic acid)

To a suspension of intermediate (IVd) (690 mg, 1.14 mmol) in DMSO (54 mL) was added a solution of lithium hydroxide (140 mg, 5.71 mmol) in water (9.0 mL). The mixture was heated at 70 ° C. for 22 hours, then cooled to RT, diluted with water (100 mL) and acidified (to pH 2) by addition of 1M hydrochloric acid (6.0 mL). The mixture was cooled in an ice bath for 15 min and the resulting precipitate was collected by filtration, washed with water (3 × 50 mL) and dried in vacuo at 40 ° C. to give intermediate (IIc) of the title compound. Was obtained as a yellow solid (617 mg, 87%); R ^t 1.91 min (method a); m / z 591 (M + H) ⁺ (ES ⁺ );

4-フルオロアニリン(IIIa)(0.85mL、9.00mmol)、トリエチルアミン(1.88mL、13.5mmol)、及びDMAP(0.11g、0.90mmol)のTHF(15mL)溶液に、4-ブロモベンゾイルクロリド(XIXa)(2.37g、10.8mmol)を添加した。反応混合物をRTで1時間保持し、その後、EtOAc(100mL)と1M塩酸(100mL)に分配した。有機相を分離し、1M塩酸(100mL)、飽和水性NaHCO₃(100mL)、及びブライン(100mL)で順次洗浄し、その後、乾燥させ、真空中で蒸発させた。粗残渣を温かいDCM(100mL)から粉砕し、混合物を還流状態で加熱すると、白色の懸濁液が得られ、これをRTに冷却させておいた。得られた沈殿物を濾過により回収すると、表題化合物の中間体(XVIIIa)が白色の固体(1.81g、65％)として得られた; R^t 2.23 分; m/z 294/296(M+H)⁺(ES⁺);

(4-Bromo-N- (4-fluorophenyl) benzamide)

To a solution of 4-fluoroaniline (IIIa) (0.85 mL, 9.00 mmol), triethylamine (1.88 mL, 13.5 mmol), and DMAP (0.11 g, 0.90 mmol) in THF (15 mL), 4-bromobenzoyl chloride (XIXa) ( 2.37 g, 10.8 mmol) was added. The reaction mixture was kept at RT for 1 h and then partitioned between EtOAc (100 mL) and 1M hydrochloric acid (100 mL). The organic phase was separated and washed sequentially with 1M hydrochloric acid (100 mL), saturated aqueous NaHCO ₃ (100 mL), and brine (100 mL), then dried and evaporated in vacuo. The crude residue was triturated from warm DCM (100 mL) and the mixture was heated at reflux to give a white suspension which was allowed to cool to RT. The resulting precipitate was collected by filtration to give the title compound intermediate (XVIIIa) as a white solid (1.81 g, 65%); R ^t 2.23 min; m / z 294/296 (M + H ) ⁺ (ES ⁺ );

tert-ブチルピペラジン-1-カルボキシレート(IXa)(4.00g、215mmol)、中間体(XVIIIa)(6.63g、22.6mmol)、RuPhos(100mg、0.215mmol)、及びRuPhos G3(180mg、0.215mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した。LiHMDS(THF中1M、75.0mL、75.0mmol)の溶液を添加し、反応混合物を70℃で5時間加熱した。RTに冷却した後、該混合物をEtOAc(150mL)と1M塩酸(150mL)に分配した。有機相を分離し、保持し、水相をEtOAc(3×150mL)で抽出した。合わせた有機物を乾燥させ、真空中で濃縮すると、茶色の固体が得られ、これをイソヘキサンとジエチルエーテル(1:1、100mL)の混合物中で粉砕した。そのようにして得られた生成物を濾過により回収し、イソヘキサンとジエチルエーテル(1:1、25mL)の混合物で洗浄し、その後、真空中、40℃で乾燥させると、表題化合物の中間体(XVIIa)が褐色の固体(6.44g、85％)として得られた; R^t 2.40分(方法a); m/z 400(M+H)⁺;

(tert-butyl 4- (4-((4-fluorophenyl) carbamoyl) phenyl) piperazine-1-carboxylate)

tert-Butyl piperazine-1-carboxylate (IXa) (4.00 g, 215 mmol), intermediate (XVIIIa) (6.63 g, 22.6 mmol), RuPhos (100 mg, 0.215 mmol), and RuPhos G3 (180 mg, 0.215 mmol) The charged flask was evacuated and refilled with nitrogen three times. A solution of LiHMDS (1M in THF, 75.0 mL, 75.0 mmol) was added and the reaction mixture was heated at 70 ° C. for 5 h. After cooling to RT, the mixture was partitioned between EtOAc (150 mL) and 1M hydrochloric acid (150 mL). The organic phase was separated and retained and the aqueous phase was extracted with EtOAc (3 × 150 mL). The combined organics were dried and concentrated in vacuo to give a brown solid that was triturated in a mixture of isohexane and diethyl ether (1: 1, 100 mL). The product so obtained was collected by filtration, washed with a mixture of isohexane and diethyl ether (1: 1, 25 mL) and then dried in vacuo at 40 ° C. to give the intermediate of the title compound ( XVIIa) was obtained as a brown solid (6.44 g, 85%); R ^t 2.40 min (method a); m / z 400 (M + H) ⁺ ;

中間体(XVIIa)(6.44g、16.1mmol)のDCM(200mL)溶液に、TFA(24.7mL、322mmol)を添加した。反応液をRTで2時間撹拌し、その後、真空中で蒸発させた。トルエン(5.0mL)を添加し、混合物を真空中で再蒸発させた。得られた油状物をDCM(90mL)とメタノール(10mL)の混合物中に取り、その後、水(50mL)と飽和水性NaHCO₃(50mL)の混合物で抽出した。有機相を分離し、保持し、水層をDCMとメタノール(9:1、3×100mL)の混合物で抽出した。合わせた有機層を乾燥させ、真空中で濃縮すると、表題化合物の中間体(XVIa)が茶色の固体(3.74g、70％)として得られた; R^t 1.02分(方法a); m/z 300(M+H)⁺;

(N- (4-Fluorophenyl) -4- (piperazin-1-yl) benzamide)

To a solution of intermediate (XVIIa) (6.44 g, 16.1 mmol) in DCM (200 mL) was added TFA (24.7 mL, 322 mmol). The reaction was stirred at RT for 2 hours and then evaporated in vacuo. Toluene (5.0 mL) was added and the mixture was re-evaporated in vacuo. The resulting oil was taken up in a mixture of DCM (90 mL) and methanol (10 mL) and then extracted with a mixture of water (50 mL) and saturated aqueous NaHCO ₃ (50 mL). The organic phase was separated and retained and the aqueous layer was extracted with a mixture of DCM and methanol (9: 1, 3 × 100 mL). The combined organic layers were dried and concentrated in vacuo to give the title compound intermediate (XVIa) as a brown solid (3.74 g, 70%); R ^t 1.02 min (method a); m / z 300 (M + H) ⁺ ;

4-ブロモ-1-メトキシ-2-メチルベンゼン(Xe)(406mg、2.02mmol)、中間体(XVIa)(550mg、1.84mmol)、RuPhos(43mg、0.092mmol)、及びRuPhos G3(77mg、0.092mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した。LiHMDS(9.2mL、THF中1M、9.2mmol)の溶液を添加し、反応混合物を70℃で8時間加熱した。RTに冷却した後、該混合物を1M水性塩酸(9.0mL)の添加によりクエンチし、その後、水(15mL)とEtOAc(15mL)に分配した。有機層を分離し、保持し、水層をEtOAc(2×15mL)で抽出した。合わせた有機物をブライン(20mL)で洗浄し、その後、乾燥させ、真空中で蒸発させた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、イソヘキサン中0〜100％EtOAc、勾配溶出)により精製すると、黄色の固体が得られた。この材料をフラッシュカラムクロマトグラフィー(SiO₂、4g、DCM中0〜10％EtOAc、勾配溶出)により再精製すると、表題化合物の中間体(XVa)がオフホワイト色の固体(83mg、11％)として得られた; R^t 2.27分(方法a); m/z 420(M+H)⁺(ES⁺);

(N- (4-Fluorophenyl) -4- (4- (4-methoxy-3-methylphenyl) piperazin-1-yl) benzamide)

4-Bromo-1-methoxy-2-methylbenzene (Xe) (406 mg, 2.02 mmol), intermediate (XVIa) (550 mg, 1.84 mmol), RuPhos (43 mg, 0.092 mmol), and RuPhos G3 (77 mg, 0.092 mmol) ) Was evacuated from the flask and refilled with nitrogen three times. A solution of LiHMDS (9.2 mL, 1M in THF, 9.2 mmol) was added and the reaction mixture was heated at 70 ° C. for 8 hours. After cooling to RT, the mixture was quenched by the addition of 1M aqueous hydrochloric acid (9.0 mL) and then partitioned between water (15 mL) and EtOAc (15 mL). The organic layer was separated and retained and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organics were washed with brine (20 mL) then dried and evaporated in vacuo. The so obtained crude product by flash column chromatography (SiO _2, 12 g, isohexane 0 to 100% EtOAc, gradient elution) to give a yellow solid. This material was purified by flash column chromatography _{(SiO 2, 4g, 0~10%} EtOAc in DCM, gradient elution) and re-purified by intermediate of the title compound (XVa) as a off-white solid (83 mg, 11%) Obtained; R ^t 2.27 min (method a); m / z 420 (M + H) ⁺ (ES ⁺ );

0℃の中間体(XVa)(83mg、0.20mmol)のDCM(5.0mL)懸濁液に、三臭化ホウ素(0.59mL、DCM中1M、0.59mmol)の溶液を添加した。反応混合物を0℃で30分間撹拌し、RTに8時間温めておき、その後、水(15mL)とDCM(10mL)に分配した。有機層を分離し、保持し、水層をDCMとMeOH(90:10、5×15mL)の混合物で抽出した。合わせた有機物を乾燥させ、真空中で蒸発させると、粗生成物が得られ、これをフラッシュカラムクロマトグラフィー(SiO₂、4.0g、DCM中0〜3％MeOH、勾配溶出)により精製すると、表題化合物の中間体(XVb)がベージュ色の固体(61mg、72％)として得られた; R^t 1.73分(方法a); m/z 406(M+H)⁺(ES⁺);

(N- (4-Fluorophenyl) -4- (4- (4-hydroxy-3-methylphenyl) piperazin-1-yl) benzamide)

To a suspension of intermediate (XVa) (83 mg, 0.20 mmol) in DCM (5.0 mL) at 0 ° C. was added a solution of boron tribromide (0.59 mL, 1 M in DCM, 0.59 mmol). The reaction mixture was stirred at 0 ° C. for 30 min and allowed to warm to RT for 8 h before being partitioned between water (15 mL) and DCM (10 mL). The organic layer was separated and retained, and the aqueous layer was extracted with a mixture of DCM and MeOH (90:10, 5 × 15 mL). The combined organics were dried and evaporated in vacuo, the crude product is obtained, by flash column chromatography _{(SiO 2, 4.0g, 0~3%} MeOH in DCM, gradient elution) to give the title Compound intermediate (XVb) was obtained as a beige solid (61 mg, 72%); R ^t 1.73 min (method a); m / z 406 (M + H) ⁺ (ES ⁺ );

4-ブロモ-2,5-ジメチルフェノール(Xf)(73.1mg、0.364mmol)、中間体(XVIa)(120mg、0.400mmol)、RuPhos(8.48mg、0.0180mmol)、及びRuPhos G3(15.2mg、0.0180mmol)を仕込んだフラスコから空気を排出し、窒素を3回再充填した。LiHMDS(THF中1M、1.46mL、1.46mmol)の溶液を添加し、反応混合物を70℃で18時間加熱した。RTに冷却した後、該混合物を1M塩酸(5.0mL)の添加によりクエンチし、その後、2M水性NaOH(10mL)で塩基性化した。水層をEtOAc(3×15mL)で抽出し、合わせた有機物を乾燥させ、真空中で蒸発させた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、24g、DCM中0〜5％メタノール(1％NH₃)、勾配溶出)により精製すると、茶色の固体が得られた。この固体をジエチルエーテル(20mL)中で粉砕し、濾過により回収し、ジエチルエーテル(10mL)で洗浄し、真空中、40℃で乾燥させると、表題化合物の中間体(XVc)がオフホワイト色の固体(72.0mg、47％)として得られた; R^t 2.19分(方法a); m/z 420(M+H)⁺(ES⁺);

(N- (4-Fluorophenyl) -4- (4- (4-hydroxy-2,5-dimethylphenyl) piperazin-1-yl) benzamide)

4-Bromo-2,5-dimethylphenol (Xf) (73.1 mg, 0.364 mmol), intermediate (XVIa) (120 mg, 0.400 mmol), RuPhos (8.48 mg, 0.0180 mmol), and RuPhos G3 (15.2 mg, 0.0180) mmol) was evacuated and refilled with nitrogen three times. A solution of LiHMDS (1M in THF, 1.46 mL, 1.46 mmol) was added and the reaction mixture was heated at 70 ° C. for 18 hours. After cooling to RT, the mixture was quenched by the addition of 1M hydrochloric acid (5.0 mL) and then basified with 2M aqueous NaOH (10 mL). The aqueous layer was extracted with EtOAc (3 × 15 mL) and the combined organics were dried and evaporated in vacuo. The so obtained crude product by flash column chromatography (SiO _2, 24g, 0~5% methanol in DCM (1% NH _3), gradient elution) to give a brown solid. This solid was triturated in diethyl ether (20 mL), collected by filtration, washed with diethyl ether (10 mL), and dried in vacuo at 40 ° C. to give the title compound intermediate (XVc) as an off-white color. Obtained as a solid (72.0 mg, 47%); R ^t 2.19 min (method a); m / z 420 (M + H) ⁺ (ES ⁺ );

0℃の中間体(IIa)(1.00g、1.70mmol)、DIPEA(1.78mL、10.2mmol)、及び塩化アンモニウム(0.454g、8.48mmol)のDMF(30mL)溶液に、HATU(1.30g、3.39mmol)を2分かけて少しずつ添加した。反応混合物をRTに2時間温め、その後、水(70mL)で希釈し、得られた固体を濾過により回収した。フィルターケーキを水(2×50mL)で洗浄し、固体を、真空中、40℃で乾燥させると、表題化合物の中間体(XXa)がオフホワイト色の粉末(850mg、83％)として得られた; R^t 2.24分(方法b); m/z 589(M+H)⁺(ES⁺);

(4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran- 3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) benzamide)

To a solution of intermediate (IIa) (1.00 g, 1.70 mmol), DIPEA (1.78 mL, 10.2 mmol), and ammonium chloride (0.454 g, 8.48 mmol) at 0 ° C. in DMF (30 mL), HATU (1.30 g, 3.39 mmol). ) Was added in portions over 2 minutes. The reaction mixture was warmed to RT for 2 h then diluted with water (70 mL) and the resulting solid was collected by filtration. The filter cake was washed with water (2 × 50 mL) and the solid was dried in vacuo at 40 ° C. to give the title compound intermediate (XXa) as an off-white powder (850 mg, 83%). R ^t 2.24 min (method b); m / z 589 (M + H) ⁺ (ES ⁺ );

中間体(IIa)(2.50g、4.24mmol)、EDCI(1.63g、8.48mmol)、及びDMAP(30mg、0.21mmol)のピリジン(30mL)懸濁液に、4-フルオロアニリン(0.41mL、4.3mmol)を添加し、反応混合物を60℃で2時間加熱し、その後、RTに冷却した。該混合物を水(60mL)で希釈し、5分間撹拌すると、固体が生じ、これを濾過により回収し、その後、水(3×10mL)及びジエチルエーテル(2×15mL)で洗浄すると、褐色の粉末が得られた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、40g、DCM中0〜3％MeOH、勾配溶出)により精製すると、表題化合物の実施例1が黄色の固体(2.47g、85％)として得られた; R^t 2.60分(方法a); m/z 683(M+H)⁺(ES⁺);

(Preparation of Compound Examples of the Invention)
Example 1: 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide)
(1.From intermediate (IIa))

To a suspension of intermediate (IIa) (2.50 g, 4.24 mmol), EDCI (1.63 g, 8.48 mmol), and DMAP (30 mg, 0.21 mmol) in pyridine (30 mL), 4-fluoroaniline (0.41 mL, 4.3 mmol). ) Was added and the reaction mixture was heated at 60 ° C. for 2 h and then cooled to RT. The mixture is diluted with water (60 mL) and stirred for 5 minutes to produce a solid that is collected by filtration and then washed with water (3 × 10 mL) and diethyl ether (2 × 15 mL) to give a brown powder was gotten. The so obtained crude product by flash column chromatography _{(SiO 2, 40g, 0~3%} MeOH in DCM, gradient elution) to afford Example 1 of the title compound as a yellow solid (2.47 g, 85%); R ^t 2.60 min (method a); m / z 683 (M + H) ⁺ (ES ⁺ );

中間体(XVc)(19mg、0.047mmol)のDMSO(1.5mL)溶液に、水性水酸化ナトリウム(1M、98μL、0.098mmol)を添加した。混合物をRTで10分間撹拌し、その後、トシレート(XIa)(APIChem製、カタログ番号: AC-8330、12.4mg、27.6mmol)のDMSO(0.5mL)溶液で処理した。反応混合物を60℃で2時間撹拌し、RTに冷却し、水(10mL)を添加した。得られた混合物をEtOAc(3×10mL)で抽出し、合わせた有機抽出物を乾燥させ、真空中で蒸発させると、茶色の油状物が得られた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、4g、DCM中0〜2％MeOH、勾配溶出)により精製すると、ベージュ色の固体(23mg)が得られた。生成物をフラッシュカラムクロマトグラフィー(SiO₂、4.0g、DCM中0〜50％EtOAc、勾配溶出)により再精製すると、表題化合物の実施例1がオフホワイト色の固体(14mg、42％)として得られた; R^t 2.60分(方法a); m/z 683(M+H)⁺(ES⁺)。 (2.From intermediate (XVc))

To a solution of intermediate (XVc) (19 mg, 0.047 mmol) in DMSO (1.5 mL) was added aqueous sodium hydroxide (1M, 98 μL, 0.098 mmol). The mixture was stirred at RT for 10 min and then treated with a solution of tosylate (XIa) (APIChem, catalog number: AC-8330, 12.4 mg, 27.6 mmol) in DMSO (0.5 mL). The reaction mixture was stirred at 60 ° C. for 2 h, cooled to RT and water (10 mL) was added. The resulting mixture was extracted with EtOAc (3 × 10 mL) and the combined organic extracts were dried and evaporated in vacuo to give a brown oil. So obtained crude product was purified by flash column chromatography _{(SiO 2, 4g, 0~2%} MeOH in DCM, gradient elution) to give a beige solid (23 mg) was obtained. The product was purified by flash column chromatography _{(SiO 2, 4.0g, 0~50%} EtOAc in DCM, gradient elution) and re-purified by, obtained in Example 1 of the title compound as an off-white solid (14 mg, 42%) R ^t 2.60 min (method a); m / z 683 (M + H) ⁺ (ES ⁺ ).

中間体(IIa)(74.0mg、0.125mmol)のDCM(1.0mL)懸濁液に、1-クロロ-N,N,2-トリメチルプロパ-1-エン-1-アミン(50.0μL、1.88mmol)を添加し、反応混合物をRTで18時間撹拌し、その後、真空中で蒸発させた。得られた褐色の固体をDCM(1.0mL)に溶解させ、2,4-ジフルオロアニリン(19.0μL、0.188mmol)のピリジン(0.5mL)溶液に添加した。反応液をRTで3時間撹拌し、その後、DCM(3.0mL)で希釈し、1M塩酸(6.0mL)の添加により、pH 2に酸性化した。混合物を相分離器に通し、有機物を真空中で蒸発させた。そのようにして得られた粗生成物をフラッシュカラムクロマトグラフィー(SiO₂、12g、イソヘキサン中50〜100％EtOAc、勾配溶出)により精製すると、表題化合物の実施例2がオフホワイト色の固体(59.0mg、66％)として得られた; R^t 2.53分(方法a); m/z 701(M+H)⁺(ES⁺);

Example 2: 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (2,4-difluorophenyl) benzamide)

To a suspension of intermediate (IIa) (74.0 mg, 0.125 mmol) in DCM (1.0 mL) was added 1-chloro-N, N, 2-trimethylprop-1-en-1-amine (50.0 μL, 1.88 mmol). Was added and the reaction mixture was stirred at RT for 18 h and then evaporated in vacuo. The resulting brown solid was dissolved in DCM (1.0 mL) and added to a solution of 2,4-difluoroaniline (19.0 μL, 0.188 mmol) in pyridine (0.5 mL). The reaction was stirred at RT for 3 h, then diluted with DCM (3.0 mL) and acidified to pH 2 by the addition of 1M hydrochloric acid (6.0 mL). The mixture was passed through a phase separator and the organics were evaporated in vacuo. So obtained crude product was purified by flash column chromatography (SiO _2, 12 g, 50 to 100% EtOAc in isohexane, gradient elution) to give Example 2 of the title compound as an off-white solid (59.0 mg, 66%); R ^t 2.53 min (method a); m / z 701 (M + H) ⁺ (ES ⁺ );

中間体(IIc)(100mg、0.169mmol)、EDCI(64.9mg、0.339mmol)、及びDMAP(1.03mg、8.47μmol)のピリジン(900μL)懸濁液に、4-フルオロアニリン(17.6μL、0.186mmol)を添加した。反応混合物をRTで18時間撹拌し、その後、水(50mL)に注ぎ入れた。得られた固体を濾過により回収し、水(10mL)及びジエチルエーテル(10mL)で洗浄し、真空中、40℃で乾燥させると、表題化合物の実施例3がオフホワイト色の粉末(94.0mg、80％)として得られた; R^t 2.25分(方法a); m/z 684(M+H)⁺(ES⁺);

Example 3: 4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide)

To a suspension of intermediate (IIc) (100 mg, 0.169 mmol), EDCI (64.9 mg, 0.339 mmol), and DMAP (1.03 mg, 8.47 μmol) in pyridine (900 μL), 4-fluoroaniline (17.6 μL, 0.186 mmol). ) Was added. The reaction mixture was stirred at RT for 18 hours and then poured into water (50 mL). The resulting solid was collected by filtration, washed with water (10 mL) and diethyl ether (10 mL) and dried in vacuo at 40 ° C. to give Example 3 of the title compound as an off-white powder (94.0 mg, 80%); R ^t 2.25 min (method a); m / z 684 (M + H) ⁺ (ES ⁺ );

中間体(IIb)(72.0mg、0.120mmol)、EDCI(45.1mg、0.235mmol)、及びDMAP(1.00mg、8.19μmol)のピリジン(1.0mL)懸濁液に、4-フルオロアニリン(11.1μL、0.118mmol)を添加し、反応混合物をRTで72時間撹拌した。該混合物を水(30mL)で希釈し、5分間撹拌した。そのようにして形成された固体を濾過により回収し、フラッシュカラムクロマトグラフィー(SiO₂、4g、DCM中0〜10％MeOH、勾配溶出)により精製すると、表題化合物の実施例4が白色の固体(36.8mg、44％)として得られた; R^t 2.59分(方法a); 694 m/z (M+H)⁺(ES⁺);

Example 4: 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -3-cyanophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide)

To a suspension of intermediate (IIb) (72.0 mg, 0.120 mmol), EDCI (45.1 mg, 0.235 mmol), and DMAP (1.00 mg, 8.19 μmol) in pyridine (1.0 mL), 4-fluoroaniline (11.1 μL, 0.118 mmol) was added and the reaction mixture was stirred at RT for 72 h. The mixture was diluted with water (30 mL) and stirred for 5 minutes. The so-formed solid was collected by filtration and purified by flash column chromatography _{(SiO 2, 4g, 0~10%} MeOH in DCM, gradient elution) to give Example 4 of the title compound as a white solid ( 36.8 mg, 44%); R ^t 2.59 min (method a); 694 m / z (M + H) ⁺ (ES ⁺ );

30℃の中間体(XVc)(70.0mg、0.167mmol)のDMSO(1.0mL)溶液に、水性NaOH(20.0μL、12.5M、0.250mmol)を添加した。30分後、トシレート(XIa)(83.0mg、0.184mmol)を添加し、反応混合物を30℃で18時間撹拌し、その後、RTに冷却し、水(15mL)に注ぎ入れた。得られた沈殿物を濾過により回収し、水(20mL)で洗浄すると、白色の固体が得られた。粗生成物を分取HPLC(方法2)により精製すると、表題化合物の実施例5が白色の固体(34.0mg、29％)として得られた; R^t 2.89分(方法a); m/z 697(M+H)⁺(ES⁺);

Example 5: 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -2,5-dimethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide)

To a solution of intermediate (XVc) (70.0 mg, 0.167 mmol) in DMSO (1.0 mL) at 30 ° C. was added aqueous NaOH (20.0 μL, 12.5 M, 0.250 mmol). After 30 minutes, tosylate (XIa) (83.0 mg, 0.184 mmol) was added and the reaction mixture was stirred at 30 ° C. for 18 hours, then cooled to RT and poured into water (15 mL). The resulting precipitate was collected by filtration and washed with water (20 mL) to give a white solid. The crude product was purified by preparative HPLC (Method 2) to give the title compound, Example 5, as a white solid (34.0 mg, 29%); R ^t 2.89 min (Method a); m / z 697 (M + H) ⁺ (ES ⁺ );

中間体(IIa)(100mg、0.17mmol)、EDCI(65mg、0.34mmol)、及びDMAP(1.04mg、8.48μmol)のピリジン(1.40mL)懸濁液に、5-フオロピリジン(fuoropyridin)-2-アミン(23.9mg、0.25mmol)を添加し、反応混合物をRTで15時間撹拌した。該混合物をDCM(8.0mL)で希釈し、1M塩酸(2.0mL)の添加により、酸性化した。その後、混合物を相分離器に通し、有機物を真空中で蒸発させた。粗生成物を分取HPLC(方法2)により精製すると、表題化合物の実施例6が白色の固体(41.0mg、34％)として得られた; R^t 2.51分(方法a); m/z 684(M+H)⁺(ES⁺);

Example 6: 4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide)

To a suspension of intermediate (IIa) (100 mg, 0.17 mmol), EDCI (65 mg, 0.34 mmol), and DMAP (1.04 mg, 8.48 μmol) in pyridine (1.40 mL), 5-fuoropyridin-2-amine (23.9 mg, 0.25 mmol) was added and the reaction mixture was stirred at RT for 15 h. The mixture was diluted with DCM (8.0 mL) and acidified by addition of 1M hydrochloric acid (2.0 mL). The mixture was then passed through a phase separator and the organics were evaporated in vacuo. The crude product was purified by preparative HPLC (Method 2) to give the title compound, Example 6, as a white solid (41.0 mg, 34%); R ^t 2.51 min (Method a); m / z 684 (M + H) ⁺ (ES ⁺ );

中間体(XXa)(100mg、0.170mmol)、6-ブロモニコチノニトリル(31.1mg、0.170mmol)、Xantphos(4.91mg、8.49μmol、5mol％)、トリス(ジベンジリデンアセトン)ジパラジウム(0)(3.89mg、4.25μmol、2.5mol％)、及び炭酸セシウム(166mg、0.510mmol)を仕込んだバイアルから空気を排出し、窒素を3回再充填した後、DMF(1.0mL)を添加した。反応液を100℃に18時間加熱し、その後、RTに冷却し、DCM(15mL)とブライン(20mL)に分配した。相を分離し、有機層を乾燥させ、真空中で濃縮すると、油性残渣が得られ、これをメタノール(6.0mL)中で粉砕した。得られた固体を濾過により単離し、ジエチルエーテル(10mL)で洗浄し、真空中、40℃で乾燥させると、粗固体が得られた。粉砕工程をメタノール(3.0mL)を用いて繰り返し、真空中で乾燥させると、表題化合物の実施例7がオフホワイト色の粉末(71.5mg、59％)として得られた; R^t 2.71分(方法b); m/z 691(M+H)⁺(ES⁺);

Example 7: 4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluoro Phenyl) tetrahydrofuran-3-yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide)

Intermediate (XXa) (100 mg, 0.170 mmol), 6-bromonicotinonitrile (31.1 mg, 0.170 mmol), Xantphos (4.91 mg, 8.49 μmol, 5 mol%), tris (dibenzylideneacetone) dipalladium (0) ( Air was evacuated from a vial charged with 3.89 mg, 4.25 μmol, 2.5 mol%) and cesium carbonate (166 mg, 0.510 mmol) and refilled with nitrogen three times before adding DMF (1.0 mL). The reaction was heated to 100 ° C. for 18 hours, then cooled to RT and partitioned between DCM (15 mL) and brine (20 mL). The phases were separated and the organic layer was dried and concentrated in vacuo to give an oily residue that was triturated in methanol (6.0 mL). The resulting solid was isolated by filtration, washed with diethyl ether (10 mL) and dried in vacuo at 40 ° C. to give a crude solid. The grinding process was repeated with methanol (3.0 mL) and dried in vacuo to give the title compound Example 7 as an off-white powder (71.5 mg, 59%); R ^t 2.71 min (method b); m / z 691 (M + H) ⁺ (ES ⁺ );

The following compound examples (Table 2) can be prepared by synthetic methods similar to the previous examples or by methods described elsewhere herein:
Table 2: Further compound examples of the invention

(Biological test: experimental method)
(Evaluation of planktonic fungal growth)
(a. Resazurin microtiter assay)
This assay was performed using a modified version of the published method (Monteiro et al., 2012). Spores of Aspergillus fumigatus (NCPF2010, Public Health England, Wiltshire) were cultured in Sabouraud dextrose agar for 3 days. Stock spore suspensions were prepared from Sabouraud dextrose agar cultures by washing with PBS-tween (10 mL; PBS containing 0.05% Tween-20, 100 U / mL penicillin, and 100 U / mL streptomycin). The number of spores was evaluated using Neubauer hemocytometer and adjusted to 10 ⁶ spores / mL with PBS. RPMI- containing 2 mM L-glutamine, 2% glucose, and 0.165 M MOPS buffered to pH 7 with filter-sterilized MOPS RPMI-1640 (50 mL; NaOH) with a working suspension of spores (10 ⁴ spores / mL) 1640). Resazurin sodium salt (100 μL of 1% solution; Sigma-Aldrich, Dorset, UK) was added to the spore suspension and mixed well. Spore suspension-resazurin mixture (100 μL / well) was added to a 384 well plate (Cat. No. 353962, BD Falcon, Oxford, UK).

At the same time, test compounds (0.5 μL DMSO solution) were added in quadruplicate to 100 μL of spore-resazurin mixture using Integra VIAFLO 96 (Intergra, Zizers, Switzerland) to provide a 0.5% final DMSO solution. For non-spore control wells, MOPS-RPMI-resazurin solution (100 μL) was added instead of the spore-resazurin mixture. Plates were covered with Breathe Easier membrane (Cat. # Z763624, Sigma-Aldrich, Dorset, UK) and incubated (35 ° C, until the fluorescence in the inoculated wells was twice that of the control wells (approximately 24 hours). 5% CO ₂ ). The fluorescence (545 nm (excitation) / 590 nm (emission), gain 800, focal height 5.5 mm) of each well was determined using a multi-scanner (Clariostar: BMG, Buckinghamshire, UK). The percent inhibition for each well was calculated and the MIC ₅₀ , MIC ₇₅ , and MIC ₉₀ values were calculated from the concentration-response curves generated for each test compound.

(b. Trace liquid dilution assay)
This assay was performed using a modified version of the method published by EUCAST (Rodriguez-Tudela et al., 2008). Sporerodextrose spores of Aspergillus fumigatus (NCPF2010, NCPF7010 (Methionine 220 mutation), NCPF7099 (Glycine G54 mutation) from Public Health England, Wiltshire; TR34 / L98H mutant from St Louis Hospital, Paris, France) Cultivated in agar for 3 days. Stock spore suspensions were prepared from Sabouraud dextrose agar cultures by washing with PBS-tween (10 mL; PBS containing 0.05% Tween-20, 100 U / mL penicillin, and 100 U / mL streptomycin). The number of spores was evaluated using a Neubauer hemocytometer and then adjusted to 10 ⁶ spores / mL with PBS. A working suspension of spores (2 × 10 ⁵ spores / mL) was filter sterilized BSA MOPS RPMI-1640 (50 mL; buffered to pH 7 with NaOH, 2 mM L-glutamine, 0.5% BSA, 2% glucose, 0.165) It was prepared in RPMI-1640) containing M MOPS. For this assay, BSA MOPS RPMI-1640 (50 μL / well) was first added to the whole of a 384 well plate (Cat # 353962, BD Falcon, Oxford, UK). Then, using Integra VIAFLO 96 (Integra, Zizers, Switzerland), the test compound (0.5 μL DMSO solution) was added in quadruplicate and mixed well using a plate mixer. Subsequently, 50 μL of working spore suspension prepared as described above was added to all wells except non-spore control wells. For non-spore control wells, BSA MOPS-RPMI solution (50 μL / well) was added instead. The plate was covered with a plastic lid and incubated for 48 hours (35 ° C., ambient air). The OD at 530 nm of each well was determined using a multi-scanner (Clariostar: BMG, Buckinghamshire, UK). The inhibition rate for each well was calculated, and the MIC ₅₀ , MIC ₇₅ , and MIC ₉₀ values were calculated from the concentration-response curves generated for each test compound.

Screening of the fungal panel was performed by Eurofins Panlabs. The guidelines for the Clinical and Laboratory Standards Institute are the micro liquid dilution method for yeast (CLSI M27-A2) (CLSI literature, 2002) and the micro liquid dilution method for filamentous fungi (CLSI M38-A). The MIC and MIC ₅₀ values of the specimens were determined according to CLSI literature (2008).

(Aspergillus fumigatus infection of bronchial epithelial cells)
BEAS2B cells were seeded in 96% plates (100 μL; 30,000 cells / well; catalog number 3596, Sigma Aldrich, Dorset, UK) in 10% FBS RPMI-1640 and then incubated for 1 day (37 ° C., 5% After CO ₂ ), the experiment was conducted. Test compound (0.5 μL DMSO solution) or vehicle (DMSO) was added to each well to give a final DMSO concentration of 0.5%. BEAS2B cells were incubated with test compounds for 1 hour (35 ° C., 5% CO ₂ ) followed by Aspergillus fumigatus (20 μL; Public Health England) conidial spore suspension (0.5 × 10 ^{5 in} 10% FBS RPMI-1640) / ml). Plates were incubated for 24 hours (35 ° C., 5% CO ₂ ). The supernatant (50 μL) was collected and transferred to a PCR plate (Catalog No. L1402-9700, Starlab, Milton Keynes, UK) and frozen until use (−20 ° C.). After thawing, the supernatant (5 μL) was diluted 1:20 by adding R7-PBS solution (95 μL; R7 vs. PBS is 1: 4; Bio-Rad Laboratories, Redmond, WA, USA). GM levels in these samples (50 μL) were measured using the Platelia GM-EIA kit (Bio-Rad Laboratories, Redmond, WA, USA). Percent inhibition was calculated for each well and IC ₅₀ values were calculated from the concentration-response curves generated for each test compound.

(Aspergillus fumigatus infection of human alveolar bilayer)
An in vitro model of human alveoli consisting of a bilayer of human alveolar epithelial cells and endothelial cells was prepared as previously described (Hope et al., 2007). This system allows administration of the test compound to the upper compartment (“atmosphere” space) and / or the lower compartment (“systemic” space). This flexibility has been exploited to study the effect of combination therapy by administering Compound Example 1 to the upper chamber and posaconazole or other antifungal agent to the lower chamber. Collected primary human pulmonary artery endothelial cells (HPAEC), and diluted EGM-2 medium (Lonza, Basel, Switzerland) in 10 ⁶ cells / mL in. The transwell was turned over and the cell suspension (100 μL / well) was applied to the bottom of each transwell. The inverted transwells were incubated for 2 hours at RT in an airflow hood, after which they were turned upside down. EGM-2 medium was added to the lower compartment (700 μL / well) and the upper compartment (100 μL / well) and the transwells were incubated for 48 hours (37 ° C., 5% CO ₂ ). Thereafter, the EGM-2 medium in the lower compartment was replaced with fresh EGM-2 medium. A549 cells were harvested and diluted to 5 × 10 ⁵ cells / mL in 10% EBM and then added to the top compartment (100 μL / well) of all transwells and the plates were incubated for 72 hours (37 ° C. , 5% CO ₂ ). Conidia of Aspergillus fumigatus (itraconazole sensitive strain NCPF2010 and itraconazole resistant strain TR34-L98H) were cultured separately in Sabouraud dextrose agar for 3 days. Prepare stock conidia suspensions of both strains from Sabouraud dextrose agar culture by washing with PBS-tween (10 mL; PBS containing 0.05% Tween-20, 100 U / mL penicillin, and 100 U / mL streptomycin) did. The number of conidia was evaluated using a Neubauer hemocytometer and adjusted to 10 ⁶ conidia / mL with PBS. A conidial spore working stock was prepared in EBM just before use (concentration of 10 ⁵ conidia / mL).

Test compound and reference compound (or neat DMSO as vehicle) in a 24-well plate for processing the lower compartment (well containing 3 μL / 600 μL of 2% FBS EBM) and a 96-well plate for processing the upper compartment Added to appropriate wells (well containing 1 μL / 200 μL of 2% FBS EBM) to provide a final DMSO concentration of 0.5%. Medium in the upper compartment was aspirated and medium containing the appropriate test compound and reference compound or vehicle was added (100 μL / well). The transwell was then transferred to a 24-well plate containing test compound and reference compound or DMSO vehicle. After 1 hour incubation (35 ° C., 5% CO ₂ ), conidia spore suspension (10 μL / well) was added to the upper compartment of each transwell. The plates were then incubated for 24 hours (35 ° C., 5% CO ₂ ). The supernatant from each compartment (5 μL / compartment) was collected and stored (−20 ° C.). After recovery of the supernatant, the medium was changed daily and all wells were treated with test compound and reference compound or DMSO for 3 days as described above. Samples were collected until fungal growth was visible in all transwells. Thereafter, the level of GM in the supernatant of the lower compartment was measured by ELISA (BioRad, CA, USA) as an indicator of Aspergillus fumigatus invasion.

(Cell survival: Resazurin assay)
BEAS2B cells were seeded in RPMI-LHC8 (RPMI-1640 and LHC8 medium combined at the same ratio) and seeded in a 384-well plate (100 μL; 3000 / well /; BD Falcon, catalog number 353962), and experimented one day later. For cell-free control wells, RPMI-LHC8 (100 μL) was added. Test compounds (0.5 μL of DMSO solution) were added using Integra VIAFLO 96 (Integra, Zizers, Switzerland) to give a final DMSO concentration of 0.5%. BEAS2B cells were incubated with each test compound for 1 day (37 ° C / 5% CO _{2 in} RPMI-LHC8). After the addition of resazurin stock solution (5 μL, 0.04%), the plates were further incubated for 4 hours (37 ° C./5% CO ₂ ). The fluorescence of each well at 545 nm (excitation) and 590 nm (emission) was determined using a multi-scanner (Clariostar: BMG Labtech). The percent loss of cell survival was calculated for each well compared to vehicle (0.5% DMSO) treatment. Where appropriate, CC ₅₀ values were calculated from concentration-response curves generated from concentration-response curves for each test compound.

(In vivo antifungal activity)
Aspergillus fumigatus (ATCC 13073 [strain: NIH 5233], American Type Culture Collection, Manassas, VA, USA) on a malt agar (Nissui Pharmaceutical, Tokyo, Japan) plate at RT (6 ± 7 ° C.) for 6-7 days. Spores were aseptically removed from the agar plate and suspended in sterile distilled water containing 0.05% Tween 80 and 0.1% agar. On the day of infection, the spore count was assessed with a hemocytometer and the inoculum was adjusted to obtain a saline concentration of 1.67 × 10 ⁸ spores mL ⁻¹ .

  To induce immunosuppression and neutropenia, A / J mice (male, 5 weeks old) were treated with hydrocortisone (Sigma H4881; 125 mg / kg, sc) 3 days, 2 days, and 1 day before infection. And cyclophosphamide (Sigma C0768; 250 mg / kg, ip) was administered 2 days before infection. On day 0, animals were infected with a spore suspension (35 μL intranasally).

  Test compounds were administered 30 minutes before infection on day 0 and thereafter on days 1, 2, and 3 (representing prophylactic treatment), or on days 1, 2, and 3 Only (representing therapeutic treatment) was administered intranasally once a day (35 μL of a 0.08-2.00 mg / mL suspension in saline). For prophylactic treatment, test compounds (0.0032 or 0.016 mg / mL suspension 35 μL in saline) were given once a day for 7 days; then 30 minutes before infection on day 0, as well as Thereafter, it was administered intranasally either on day 1, 2, and 3 after infection, or on day 0 only. The effects of these treatment paradigms are limited to treatment 1 day and 30 minutes before inoculation, then 1 day, 2 days and 3 days after infection; or only 1 day and 30 minutes before infection. It was compared with that obtained when further reduced. Animal weights were monitored daily and those that showed a ≧ 20% decrease compared to their day 0 body weight were selected.

  Six hours after the last dose, the animals were anesthetized and the trachea was cannulated and BALF was collected. The total number of alveolar cells was determined using a hemocytometer, and the numbers of alveolar macrophages and neutrophils were determined as previously reported (Kimura et al., 2013), respectively, with anti-mouse MOMA2-FITC. FACS analysis (EPICS® ALTRA II, Beckman Coulter, Fullerton, CA, USA) using (macrophages) or anti-mouse 7/4 (neutrophils). Quantikine® mouse IFN-γ, IL-17, IL-6, or TNF-α ELISA kit for levels of IFN-γ and IL-17 in BALF and IL-6 and TNFα in serum, respectively (R & D systems, Minneapolis, MN, USA). The oxidative stress marker MDA was assayed using the OxiSelect® TBARS assay kit (MDA Quantitation; Cell Biolabs, San Diego, Calif., USA). Serum Aspergillus GM was determined using the Platelia GM-EIA kit (Bio-Rad Laboratories, Redmond, WA, USA). Cut-off index was calculated by the formula: Cut-off index = OD in sample / OD in cut-off control provided in kit. For the tissue fungal challenge assay, 100 mg of lung tissue was aseptically removed and placed in sterile distilled water and homogenized in 0.2 mL of 0.1% agar. Serially diluted lung homogenates were plated on malt agar plates (50 μL / plate) and incubated at 24 ± 1 ° C. for 72-96 hours. A. fumigatus colonies on each plate were counted and the fungal titer was presented as CFU per gram of lung tissue.

Hydrocortisone (Sigma H4881; 125 mg / kg, sc) was administered once a day for 3 days before infection, and cyclophosphamide (Sigma C0768; 250 mg / kg, ip) was administered for 2 days before infection. In addition, the effect of combined treatment of Compound Example (I) administered intranasally and posaconazole administered orally using severely immunosuppressed neutropenic A / J mice (male, 5 weeks old) Evaluated. On day 0, animals were infected intranasally with 35 μL of a spore suspension of Aspergillus fumigatus (ATCC 13073 [strain: NIH 5233]) (1.67 × 10 ⁸ spores / mL in saline). Compound Example (I), prepared as a suspension in isotonic saline (0.4 mg / mL), was administered by intranasal injection (35 μL / mouse) once daily on days 1-6 after infection. Administered. Posaconazole (1 mg / kg) was orally administered once a day, 1-6 days after infection. Body weight and survival were monitored daily until day 7.

表脚注: 1.レサズリンマイクロタイターアッセイ; 2.気管支上皮細胞; 3. n＝3〜5; (Summary of screening results)
The compounds of the present invention disclosed herein exhibit potent inhibitory activity against bronchial epithelial cell infection by azole-sensitive Aspergillus fumigatus and, when tested, fungal growth as assessed by the resazurin assay (Table 3). ). With one exception, incubation with the compounds of the invention had no or little effect on the survival of BEAS2B bronchial epithelial cells at concentrations up to at least 10 μM. In particular, in these assay systems, Compound Example 1 showed significantly greater potency than voriconazole and amphotericin B, and potency similar to posaconazole.
Table 3. Effects of treatment with voriconazole, posaconazole, amphotericin B, and compound examples of the invention on Aspergillus fumigatus (NCPF2010) floating fungal growth, on fungal infection of BEAS2B bronchial epithelial cells and on BEAS2B cell survival

Table footnotes: 1. Resazurin microtiter assay; 2. Bronchial epithelial cells; 3. n = 3-5;

表脚注: 1.微量液体希釈アッセイ、n＝3 Furthermore, the compounds of the present invention show potent inhibitory activity against planktonic fungal growth when evaluated in a microfluidic dilution assay (Table 4). In this assay, the compounds of the invention generally have significantly greater potency than posaconazole, voriconazole, and amphotericin B against posaconazole resistant strains (NCPF7099, NCPF7100, and TR34 / L98H) and posaconazole sensitive strains (NCPF2010). Indicated.
Table 4 Effect of treatment with voriconazole, posaconazole, amphotericin B, and compound examples of the present invention on planktonic fungal growth of Aspergillus fumigatus isolates

Table footnote: 1. Microfluid dilution assay, n = 3

表脚注: MIC₅₀/MIC₁₀₀＝目視検査(CLSI)による真菌成長の50％及び100％阻害に必要とされる濃度 The effect of Compound Example 1 on the growth of a wide range of fungal pathogens was evaluated using the CLSI microfluid dilution method. Compound Example 1 is a potent inhibitor of the growth of Rhizopus oryzae, Cryptococcus neoformans, Ketomium globosum, Penicillium chrysogenum, and S. aureus, and some Candida species (Table 5). I found out.
Table 5 Effect of Compound Example 1 on the growth of various fungal species

Table footnote: MIC ₅₀ / MIC ₁₀₀ = concentration required for 50% and 100% inhibition of fungal growth by visual inspection (CLSI)

表脚注: 1. 0.1μg/mLで投与した; 2. 0.01μg/mLで投与した; DFB₅₀:対照の50％の真菌量に達するのにかかる日数 Monotherapy with either Compound Example 1 (in the upper chamber, 0.1 μg / mL) or posaconazole (in the lower chamber, 0.01 μg / mL) inhibited GM production in the human alveolar bilayer on day 1. . However, the inhibitory effect of these treatments was subsequently lost rapidly (Table 6). In contrast, the combination treatment of Compound Example 1 and posaconazole showed sustained inhibition of entry after infection. As a result, the DFB ₅₀ for the combination treatment was 5.48 days, much longer than the value for either compound alone. This synergistic or additive effect of combination therapy was also confirmed when treatment with Compound Example 1 was combined with treatment with intraconazole, voriconazole, or caspofungin (results not shown).
Table 6 Effect of Compound Example 1, posaconazole, and treatment combination on Aspergillus fumigatus (NCPF2010) invasion into the lower chamber of human alveolar bilayer (transwell)

Table footnotes: 1. administered at 0.1 μg / mL; 2. administered at 0.01 μg / mL; DFB ₅₀ : days taken to reach 50% fungal load of control

表脚注: 1. 1μg/mLで投与した; 2. 0.1μg/mLで投与した; DFB₅₀:対照の50％の真菌量に達するのにかかる日数 Furthermore, this combination treatment was tested in bilayers infected with Aspergillus fumigatus azole resistant strain: TR34-L98H (Table 7). Monotherapy with Compound Example 1 (1 μg / mL) in the upper chamber or posaconazole (0.1 μg / mL) in the lower chamber showed limited benefit. In contrast, the combination of Compound Example 1 and posaconazole showed a significant inhibitory effect on fungal invasion into the lower chamber. The beneficial effect of the combination treatment was observed on day 1 after infection but disappeared after day 2.
Table 7 Effect of Compound Example 1, Posaconazole, and treatment combination on Aspergillus fumigatus (TR34-L98H strain) invasion into the lower chamber of the alveolar bilayer cell system (Transwell)

Table footnotes: 1. dosed at 1 μg / mL; 2. dosed at 0.1 μg / mL; DFB ₅₀ : days taken to reach 50% fungal load of control

表脚注: 1. 0.4mg/mLで鼻腔内に投与した; 2. 0日目の動物体重と比較した％体重減少 In direct comparison, Compound Example 1 was measured over 3 days when administered intranasally (prophylactic treatment) to immunodeficient neutropenic mice on day 0 and days 1-3 after inoculation. It showed a better effect than posaconazole on reducing weight loss caused by Aspergillus fumigatus infection (Table 8).
Table 8: Comparison of effects of Compound Example 1 and posaconazole treatment on weight loss in immunodeficient neutropenic mice caused by Aspergillus fumigatus infection

Table footnotes: 1. 0.4 mg / mL given intranasally; 2.% body weight loss compared to day 0 animal weight

表脚注:真菌負荷のデータは、平均±平均の標準誤差として示されている(SEM; n＝5〜6)。
表10:アスペルギルス・フミガーツスに感染した免疫不全好中球減少症マウスの肺における真菌負荷並びにBALF及び血清におけるガラクトマンナン濃度に対するポサコナゾール及び化合物実施例1による予防的処置のID₅₀値

Furthermore, prophylactic and therapeutic treatment with Compound Example 1 showed a better effect than posaconazole on fungal load in the lung after infection and on GM concentrations in both BALF and serum. Data for this compound used in prophylactic and therapeutic dosing regimens is shown in Table 9 and Figures 1, 2, and 3 (ID ₅₀ values are shown in Table 10).
Table 9: Effect of prophylactic and therapeutic treatment with Compound Example 1 on CFU and BALF and serum galactomannan concentrations in lungs of immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnote: Fungal burden data are presented as mean ± standard error of the mean (SEM; n = 5-6).
Table 10: ID ₅₀ values of prophylactic treatment with posaconazole and compound Example 1 on fungal load in lungs and galactomannan concentrations in BALF and serum of immunodeficient neutropenic mice infected with Aspergillus fumigatus

表12:アスペルギルス・フミガーツスに感染した免疫不全好中球減少症マウスの肺における真菌負荷及びBALFと血清の両方におけるガラクトマンナン濃度に対する試験化合物による治療的処置(1日目、2日目、及び3日目)の効果

Furthermore, prophylactic or therapeutic treatment with certain other compounds of the invention showed a better effect than posaconazole on fungal burden in the lung after infection and on GM concentrations in both BALF and serum. . Data from these studies are shown below (Tables 11 and 12).
Table 11: Prophylactic treatment with test compounds against fungal load in the lungs of lungs of immunodeficient neutropenic mice infected with Aspergillus fumigatus and galactomannan concentrations in BALF and serum (Day 0, Day 1, Day 2, And 3rd day)

Table 12: Therapeutic treatment with test compounds for fungal load in the lung and galactomannan concentrations in both BALF and serum in the lungs of immunodeficient neutropenic mice infected with Aspergillus fumigatus (Days 1, 2, and 3) (Day) effect

表脚注:細胞数のデータは、平均±平均の標準誤差(SEM)として示されている。N＝5〜6。
表14:アスペルギルス・フミガーツスに感染した免疫不全好中球減少症マウスのBALFにおけるIL-17、IFNγ、及びMDAレベルに対するポサコナゾール及び化合物実施例1による予防的処置のID₅₀値

Prophylactic treatment with Compound Example 1 also inhibited inflammatory cell accumulation in BALF in a manner similar to posaconazole (Table 13). Furthermore, the prophylactic treatment with Compound Example 1 showed an inhibitory effect superior to posaconazole on IL-17, IFNγ, and MDA concentrations in BALF. Comparative ID ₅₀ values for Compound Example 1 and posaconazole in independent experiments are shown in Table 14.
Table 13: Effect of prophylactic and therapeutic treatment with Compound Example 1 on the accumulation of macrophages and neutrophils in BALF of immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnote: Cell number data are presented as mean ± standard error of the mean (SEM). N = 5-6.
Table 14: ID ₅₀ values of prophylactic treatment with posaconazole and compound Example 1 on IL-17, IFNγ, and MDA levels in BALF of immunodeficient neutropenic mice infected with Aspergillus fumigatus

表脚注:バイオマーカー濃度のデータは、平均±平均の標準誤差(SEM)として示されている、N＝5〜6。
表16:アスペルギルス・フミガーツスに感染した免疫不全好中球減少症マウスの血清におけるIL-6及びTNFαレベルに対する化合物実施例1による予防的及び治療的処置の効果

表脚注:バイオマーカー濃度のデータは、平均±平均の標準誤差(SEM)として示されている。N＝5〜6。 In addition, data showing the effect of Compound Example 1 on the levels of IFNγ, IL-17, and MDA in BALF when administered either prophylactically or therapeutically are shown in Table 15, The effects on IL-6 and TNFα are shown in Table 16.
Table 15: Effect of prophylactic and therapeutic treatment with Compound Example 1 on IFNγ, IL-17, and MDA levels in BALF of immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnote: Biomarker concentration data are shown as mean ± standard error of the mean (SEM), N = 5-6.
Table 16: Effect of prophylactic and therapeutic treatment with Compound Example 1 on IL-6 and TNFα levels in sera of immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnote: Biomarker concentration data are presented as mean ± standard error of the mean (SEM). N = 5-6.

  Therapeutic treatment with Compound Example 1 was also found to maintain potent inhibition of pulmonary fungal burden, serum galactomannan levels, and BALF cytokine levels in immunodeficient neutropenic mice infected with Aspergillus fumigatus ( Tables 8, 9, 10, and 13; and FIGS. 1, 2, and 3).

表脚注: 1. N値は、全ての薬物処置群について6である; 2. N値は、ビヒクル処置群について5である; 3.真菌負荷及びGMレベルのデータは、ビヒクルに対する、平均±平均の標準誤差及び阻害率として示されている。 The effect of prolonged prophylactic administration of Compound Example 1 on biomarkers in immunodeficient neutropenic mice infected with Aspergillus fumigatus was also evaluated. Prolonged prophylaxis with the compound of Example 1 was found to inhibit fungal burden in the lung and GM concentrations in both BALF and serum at doses 25-fold lower than those used in previous biomarker studies (Table 17). Furthermore, this data suggests an accumulation of antifungal effects in the lungs on repeated administration, as 7 days of prevention resulted in greater antifungal effects than the additional 1 day of preventive treatment. The sustained action of the compound in the lungs shows that treatment on days -7 to 0 produces a better antifungal effect on day 3 than that obtained from treatment on days -1 and 0 only. It is suggested by the discovery that. Nevertheless, this shortened dosing protocol was still protective.
Table 17: Effect of prolonged prophylactic administration of Compound Example 1 on fungal burden (CFU) in lungs and GM concentrations in BALF and serum of immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnotes: 1. N value is 6 for all drug treatment groups; 2. N value is 5 for vehicle treatment group; 3. Fungal burden and GM level data are mean ± mean for vehicle The standard error and the inhibition rate are shown.

表脚注:群当たりN＝8。 The effect on survival of the combination of locally administered compound Example 1 treatment with oral posaconazole was evaluated in severely immunodeficient neutropenic mice after inoculation with Aspergillus fumigatus. Monotherapy with Compound Example 1 (administered intranasally, 0.4 mg / mL) or posaconazole (administered orally, 1.0 mg / kg) showed very limited therapeutic benefit. In contrast, the combination of Compound Example 1 and posaconazole showed a significant increase in survival time after infection (Table 18).
Table 18 Effects of Compound Example 1 and posaconazole as monotherapy and in combination on survival in severely immunodeficient neutropenic mice infected with Aspergillus fumigatus

Table footnote: N = 8 per group.

(In vivo pharmacokinetics)
It is common for pulmonary therapeutics to be collected at various times after administration to characterize the resulting systemic exposure to the administered compound when administered to the lungs of an animal, such as a mouse. Is the procedure used for The compounds of the present invention can be tested in such in vivo systems.

(Summary of biological profiles of compound examples of the present invention)
The compound examples of the invention disclosed herein have been found to be potent inhibitors of bronchial epithelial cell infection and buoyant growth by Aspergillus fumigatus. The compounds of the present invention also inhibited the growth of posaconazole-resistant and voriconazole-resistant Aspergillus fumigatus isolates and shared in common greater potency than posaconazole, voriconazole, and intraconazole against these strains. In vivo, in immunodeficient neutropenic mice infected with Aspergillus fumigatus, the compounds of the present invention, whether administered prophylactically or therapeutically, are potent in Aspergillus fumigatus infection. Showed inhibition. In particular, Compound Example 1, whether administered prophylactically or therapeutically, showed potent inhibition of the pulmonary immune response associated with Aspergillus fumigatus infection. Furthermore, Compound Example 1 was extremely effective in reducing infection-dependent weight loss. These inhibitory effects were superior to those of posaconazole. Significantly, the beneficial antifungal effects of the compound of Example 1 and the compounds of the invention in general are observed in both prophylactic and therapeutic dosage situations.

(Reference)

  Throughout this specification and the following claims, unless the context requires otherwise, the word `` comprise '' and variations such as `` comprises '' and `` comprising '' It is understood that it is meant to encompass the stated integer, step, group of integers, or group of steps, and does not exclude any other integer, step, group of integers, or group of steps. It will be.

Claims (54)

A compound of formula (I): or a pharmaceutically acceptable salt thereof.

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
X represents CH or N;
Y represents CH or N;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl. ) R is, H, F, CN, OCHF 2, OCF 3, SO 2 NH 2, SO 2 NHMe, SO 2 NMe 2, or Cl, especially, represents F or CN, according to claim 1, wherein formula (I) Compound.   3. A compound of formula (I) according to claim 2, wherein R represents F. The compound of formula (I) according to any one of claims 1 to 3, wherein R ^1a represents H, F or Cl, especially H. A compound of formula (I) according to claim 4, wherein R ^1b represents H or F, in particular H. R ² is, H, Me, or an OMe, compounds of formula (I) according to any one of claims 1 to 5. R ² represents H, compound of formula (I) according to claim 6, wherein. R ³ is, Me, CN, Cl, represents a CH ₂ OH, or F, compounds of formula (I) according to any one of claims 1 to 7. R ³ represents Me, compounds of formula (I) according to claim 8. R ⁴ is, H, or Me, especially, represents H, compound of formula (I) according to any one of claims 1 to 9.   11. A compound of formula (I) according to any one of claims 1 to 10, wherein X represents CH or N, especially CH. The formula (1) according to any one of claims 1 to 11, wherein Y represents CH (this carbon atom may be optionally substituted by R, R ^1a , or R ^1b , for example by F). Compound of I).   12. A compound of formula (I) according to any one of claims 1 to 11, wherein Y represents N. R ^1a and R ^1b each represent H and R represents F, CN, OCHF ₂ , OCF ₃ , SO ₂ NH ₂ , SO ₂ NHMe, SO ₂ NMe ₂ , or Cl, especially R is 14. A compound of formula (I) according to any one of claims 1 to 13 representing F or CN. ^15. A compound of formula (I) according to claim 14, wherein R ^1a and R ^1b each represent H and R represents F. R ^1b represents H, R ^1a represents F, R is, F or OCHF _2, among others, represents a F, compounds of formula (I) according to any one of claims 1 to 13. R ^1b is, represents H, R ^1a represents Cl, R represents F, compounds of formula (I) according to any one of claims 1 to 13. R ⁴ represents H, R ³ is, represents Me, CN, Cl, CH ₂ OH, or F, especially, R ⁴ represents H, R ³ represents Me, claim 1 to 17 A compound of formula (I) according to claim 1. R ⁴ represents Me, R ³ represents Me, 18. Compounds of formula (I) according. 20. Y represents CH, R ^1a and R ^1b each represent H and R is in the 3rd or 4th position, especially in the 4th position, according to any one of claims 1-12 or 14-19. A compound of formula (I) Y represents CH, R ^1b represents H, and R and R ^1a are in positions 2, 4, 3, 4, 2, 5, or 3, 5, especially 2, 4 20. The compound of formula (I) according to any one of Items 1-12 or 14-19. Any one of claims 1-11 or 13-19, wherein Y represents N, R ^1a and R ^1b each represent H, and R is in the 4th, 5th, or 6th position, especially the 5th position. A compound of formula (I) according to claim 1. 20. A compound of formula (I) according to any one of claims 1-12 or 14-19, wherein Y represents CH and R, ^R1a and ^R1b are in the 2-position, 4-position and 6-position. . R ² is in the position ortho to the nitrogen of the piperazinyl substituent, a compound of formula (I) according to any one of claims 1 to 23. R ² is in the meta position to the nitrogen of the piperazinyl substituent, a compound of formula (I) according to any one of claims 1 to 23. R ⁴ is in the position ortho to the oxygen of the ether substituent, the compounds of formula (I) according to any one of claims 1 to 25.   Aromatic moieties containing Y are 4-fluorophenyl, 3-fluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 4-cyanophenyl , 3-cyanophenyl, 4-difluoromethoxyphenyl, 4- (trifluoromethoxy) phenyl, 4-sulfamoylphenyl, 4- (N-methylsulfamoyl) phenyl, 4- (NN-dimethylsulfamoyl) Phenyl, 4-cyano-2-fluorophenyl, 4- (difluoromethoxy) -3-fluorophenyl, 4-chloro-2-fluorophenyl, 4-chloro-3-fluorophenyl, 2,4,6-trifluorophenyl 5-fluoropyridin-2-yl, 5-cyanopyridin-2-yl, 6-cyanopyridin-2-yl, 4-cyanopyridin-2-yl, 5- (trifluoromethoxy) pyridin-2-yl, Or 5-chloropyridin-2-yl, especially 4-fluoro 27. A compound of formula (I) according to any one of claims 1 to 26, which represents fluorophenyl or 4-cyanophenyl, more preferably 4-fluorophenyl.   28. A compound of formula (I) according to claim 27, wherein said aromatic moiety comprising Y represents 4-fluorophenyl or 4-cyanophenyl, more preferably 4-fluorophenyl. The following:
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (2,4-difluorophenyl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-cyanophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -2,5-dimethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (3-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-chlorophenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3,5-dimethylphenyl) piperazin-1-yl) -N- (4-cyanophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-difluoromethoxyphenyl) benzamide;
4- (4- (5-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (trifluoromethoxy) phenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3,5-dimethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-hydroxymethylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-chlorophenyl) piperazin-1-yl) -N- (4-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-sulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (N-methylsulfamoyl) phenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (4- (N, N-dimethylsulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (4-sulfamoylphenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyano-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-fluorophenyl) piperazin-1-yl) -N- (4-cyano-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4- (difluoromethoxy) -3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (2,5-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (3,4-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (3,5-difluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-chloro-2-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-chloro-3-fluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (2,4,6-trifluorophenyl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -3-methylbenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -2-methylbenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -3-methoxybenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-fluorophenyl) -2-methoxybenzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 3-methylphenyl) piperazin-1-yl) -N- (6-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (4-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-hydroxymethylphenyl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5- (trifluoromethoxy) pyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-fluorophenyl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -3-methylphenyl) piperazin-1-yl) -N- (5-chloropyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) 6-methylpyridin-2-yl) piperazin-1-yl) -N- (5-cyanopyridin-2-yl) benzamide;
4- (4- (4-(((3R, 5R) -5-((1H-1,2,4-triazol-1-yl) methyl) -5- (2,4-difluorophenyl) tetrahydrofuran-3 -Yl) methoxy) -6-methylpyridin-2-yl) piperazin-1-yl) -N- (5-fluoropyridin-2-yl) benzamide and pharmaceutically acceptable salts thereof, Item 1. The compound of formula (I) according to item 1.   30. A compound according to any one of claims 1 to 29 for use as a medicament.   30. A compound according to any one of claims 1 to 29 for use in the treatment of mycosis or for use in the prevention or treatment of diseases associated with mycosis.   30. Use of a compound according to any one of claims 1 to 29 in the manufacture of a medicament for the treatment of mycosis or for the prevention or treatment of diseases associated with mycosis.   30. A method of treating a subject having mycosis, comprising administering to the subject an effective amount of a compound according to any one of claims 1-29.   30. A method for the prevention or treatment of a disease associated with mycosis in a subject comprising administering to the subject an effective amount of a compound according to any one of claims 1-29.   35. A compound, use or method for use according to any one of claims 31 to 34, wherein the mycosis is caused by an Aspergillus species.   34. A compound, use, or method for use according to claim 33, wherein the species of the genus Aspergillus is Aspergillus fumigatus or Aspergillus pullulans, especially Aspergillus fumigatus.   36. A compound, use, or method for use according to claim 35, wherein the Aspergillus species is an azole-resistant strain of Aspergillus fumigatus.   The mycosis is a Candida species such as Candida albicans or Candida glabrata, Rhizopus species such as Rhizopus oryzae, Cryptococcus species such as Cryptococcus neoformans, Ketmium species For use according to any one of claims 31 to 34, caused by, for example, a genus Ketomium, a species of the genus Penicillium, for example a species of Penicillium chrysogenum, or a species of the genus Trichophyton, Compound, use, or method.   30. A compound according to any one of claims 1 to 29 for use as a medicament in combination with a second or further active ingredient.   30. A pharmaceutical composition comprising a compound according to any one of claims 1 to 29, optionally in combination with one or more pharmaceutically acceptable diluents or carriers.   41. A pharmaceutical composition according to claim 40, comprising a second or further active ingredient.   Said second or further active ingredient is an antifungal agent (e.g. voriconazole or posaconazole), amphotericin B, echinocandin (e.g. caspofungin), and an inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA reductase (e.g. 42. A compound for use according to claim 39 or a pharmaceutical composition according to claim 41, selected from lovastatin, pravastatin or fluvastatin).   Said second or further active ingredient is candicidin, philippines, hamycin, natamycin, nystatin, rimocidin, bifonazole, butconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxyconazole, Sertaconazole, sulconazole, thioconazole, albaconazole, efinaconazole, epoxiconazole, fluconazole, isabconazole, itraconazole, propiconazole, rubconazole, terconazole, abafungin, amorolfine, butenafine, naphthifine, terbinafine, fanfangin Benzoic acid, ciclopirox, flucytosine (5-fluorocytosine 42. A compound for use according to claim 39 or a pharmaceutical composition according to claim 41, selected from: syn), griseofulvin, tolnaftate and undecylenic acid. A compound of formula (II): or a salt thereof.

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ^a represents H; and
X represents CH or N. ) A compound of formula (IV): or a salt thereof.

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ^a represents C _1-5 alkyl; and
X represents CH or N. ) Compound of formula (V): or salt thereof

(Where:
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl; and
X represents CH or N. ) A compound of formula (VII): or a salt thereof.

(Where:
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ^b represents a nitrogen protecting group such as Boc; and
X represents CH or N. ) A compound of formula (VIII): or a salt thereof.

(Where:
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl; and
X represents CH or N. ) A compound of formula (XIII): or a salt thereof.

(Where:
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ^a represents C _1-5 alkyl; and
X represents CH or N. ) A compound of formula (XV): or a salt thereof.

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl;
X represents CH or N; and
Y represents CH or N. ) Compound of formula (XX): or a salt thereof.

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl; and
X represents CH or N. ) A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 29, comprising a compound of formula (II): or an activated derivative thereof; salt

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ^a represents hydrogen; and
X represents CH or N);
A compound of formula (III): or a salt thereof

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl; and
Y represents CH or N)
Reacting with said method. A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 29, comprising a compound of formula (XV): or a salt thereof

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo;
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl;
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl;
X represents CH or N; and
Y represents CH or N)
A compound of formula (XI): or a salt thereof

(Where:
Z represents a leaving group such as p-tolyl SO ₂ O)
Reacting with said method. A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 29, comprising a compound of formula (XX): or a salt thereof

(Where:
R ² represents hydrogen, halo, cyano, C _1-4 alkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy;
R ³ represents halo, cyano, C _1-4 alkyl, or C _1-4 hydroxyalkyl;
R ⁴ represents hydrogen or C _1-4 alkyl; and
X represents CH or N)
A compound of formula (XXI): or a salt thereof

(Where:
R represents hydrogen, halo, cyano, C _1-4 haloalkyl, C _1-4 haloalkoxy, or SO ₂ NR ⁵ R ⁶ ;
R ^1a and R ^1b independently represent hydrogen or halo; and
R ⁵ and R ⁶ independently represent hydrogen or C _1-4 alkyl)
Reacting with said method.

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